WO2024088183A1

WO2024088183A1 - Anticoronviral compounds and compositions and uses thereof

Info

Publication number: WO2024088183A1
Application number: PCT/CN2023/125810
Authority: WO
Inventors: Gongxin HE; Kai Hou; Hao Wu; Xiubo TANG; Wenyuan Fan
Original assignee: Shanghai Curegene Pharmaceutical Co., Ltd.
Priority date: 2022-10-23
Filing date: 2023-10-21
Publication date: 2024-05-02

Abstract

Provided herein are compounds which exhibit activity in the inhibition of RNA polymerase as well as pharmaceutical compositions comprising these compounds and methods of treatment of viral infections by administration of these compounds or the pharmaceutical compositions.

Description

ANTICORONVIRAL COMPOUNDS AND COMPOSITIONS AND USES THEREOF

FIELD OF THE DISCLOSURE

The present disclosure generally relates to compounds which exhibit activity in the inhibition of RNA polymerase as well as pharmaceutical compositions comprising these compounds and methods of treatment by administration of these compounds or the pharmaceutical compositions comprising the same.

BACKGROUND OF THE DISCLOSURE

Over the past two decades, multiple varieties of viruses that can cause life-threatening disease in humans and animals have been identified, namely, influenza, Respiratory syncytial virus (RSV) , parainfluenza, severe acute respiratory syndrome coronavirus (SARS-CoV) , Middle East Respiratory Syndrome coronavirus (MERS-CoV) , Ebola virus etc. Amongst, the global pandemic caused by SARS-CoV-2 has resulted in severe respiratory illness throughout the world, wherein severe cases progress to pneumonia and multiorgan failure, which have led to millions of deaths worldwide. Coronaviruses are highly susceptible to mutate into prevalent variants. Although various vaccines have been approved for use since the outbreak of SARS-CoV-2, the vaccinated people are still under risk associated with the emergence of immune escape mutants. Therefore, it is important to develop anti-viral drug to combat existing and emerging coronaviruses.

Coronaviruses are positive-strand RNA viruses encoding 16 nonstructural proteins (nsp1 to nsp16) . A number of the nonstructural proteins coalesce to form a multi-protein replicase-transcriptase complex (RTC) . The main replicase-transriptase protein is the RNA-dependent RNA polymerase (RdRp) , which is directly involved in the replication and transcription of RNA from an RNA strand. Besides, RdRps are highly conserved throughout virants and viruses, making it a desirable drug target for managing various RNA-viral infections.

Remdesivir, a monophosphoramidate prodrug of the nucleoside GS-441524, originally developed to treat Ebola virus infections, inhibits the RdRp of SARS-CoV-2. It was the first antiviral approved or authorized for emergency use to treat COVID-19 in several countries. Remdesivir improves clinical outcomes in patients hospitalized with moderate-to-severe disease and it prevents disease progression in outpatients (Beigel J.H., et al. Remdesivir for the treatment of covid-19 -final report. N. Engl. J. Med. 2020; 383 (19) : 1813-1826.; Gottlieb R.L., et al. Early remdesivir to prevent progression to severe covid-19 in outpatients. N. Engl. J. Med. 2021) . Vangeel et al. also proved that Remdesivir is effective against different variants of concern of SARS-CoV-2 (Vangeel L., et al. Remdesivir, Molnupiravir and Nirmatrelvir remain active against SARS-CoV-2 Omicron and other variants of concern. Antiviral Res. 2022 Feb; 198: 105252. ) . However, Remdesivir has several drawbacks, such as, limited to intravaneous administration, short plasma half-life, uncorrlated plasma exposure and clinical efficacy, low devlievery in lung (Sun D. Remdesivir for treatment of COVID-19: combination of pulmonary and IV administration may offer additional benefit. AAPS J. 2020; 22: 77. ) .

Accordingly, there is a need in the art to develop improved compounds which exhibit inhibitory activity against RNA polymerase, in particular, an RNA polymerase inhibitor with good oral and pulmonary bioavailability, and high conversion rate to its monophosphate and triphosphate.

SUMMARY OF THE DISCLOSURE

The present disclosure provides compounds which are capable of inhibiting RNA polymerase, the pharmaceutical compositions comprising these compounds and methods for the use of such compounds or pharmaceutical compositions for treatment of viral infections.

In one aspect, the present disclosure provides a compound having Formula (I) :

or a pharmaceutically acceptable salt thereof,

wherein

Base is a naturally occurring or modified pyrimidine base or purine base;

R¹ is selected from the group consisting of hydrogen, halogen, hydroxyl, cyano, azido, alkyl, alkenyl, alkynyl, haloalkyl, -OR^a, -NO₂, -N (R^a) ₂, -C (O) N (R^a) ₂, -C (O) R^a, -OC (O) R^a, -C (O) OR^a, -S (O) R^a, -S (O) ₂R^a, -S (O) OR^a, -S (O) ₂ (OR^a) , and -S (O) ₂N (R^a) ₂;

each of R²¹, R²², R³¹, R³², and R⁴ is independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, cyano, azido, alkyl, alkenyl, alkynyl, haloalkyl, -OR^a, -NO₂, -N (R^a) ₂, -C (O) N (R^a) ₂, -C (O) R^a, -OC (O) R^a, -C (O) OR^a, -S (O) R^a, and -S (O) ₂R^a;

Q is CH₂, CHD or CD₂;

each M is independently selected from hydrogen, metal ion, -NH₄, or protonated organic amine;

L is selected from hydrogen or -C (R⁵) ₂-L¹-L²;

each R⁵ is selected from hydrogen, deuterium, or alkyl optionally substituted with one or more groups independently selected from halogen, hydroxyl, cyano, cycloalkyl, heterocyclyl, aryl or heteroaryl;

L¹ is selected from -OC (O) -*, -OC (O) O-*, -OC (O) N (R^a) -*, -N (R^a) C (O) -*, -N (R^a) C (O) O-*, orwherein *end of L¹ is connected to L²;

L² is selected from hydrogen, R⁶, wherein *end of L² is connected to L¹;

R⁶ is selected from C_9-26 alkyl, C_9-26 alkenyl, C_9-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, R^aOCH₂ (CH₂OCH₂) _nCH₂-, wherein the alkyl, alkenyl or alkynyl is optionally substituted with one or more R^b;

R⁷ is selected from C_6-26 alkyl, C_6-26 alkenyl, C_6-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-, each of which is optionally substituted with one or more R^b;

each of R⁸ and R⁹ is independently selected from C_1-26 alkyl, C_2-26 alkenyl, C_2-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-, each of which is optionally substituted with one or more R^b;

provided that when R⁸ is methyl and R⁹ is hydrogen, then R⁷ is selected from C_7-26 alkyl, C_7-26 alkenyl, C_7-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-;

each R^a is independently selected from hydrogen, halogen or alkyl;

each R^b is independently selected from halogen, hydroxyl, cyano, amino or alkyl;

m is 0, 1, 2, or 3; and

each n, p, or q is independently an integer between 0-20.

In a further aspect, there is provided a compound having a Formula (Ia) :

or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure provides a pharmaceutical composition comprising the compound of the present disclosure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In a further aspect, the present disclosure provides a method for treating a viral infection in a patient in need thereof, comprising administering an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure to the subject.

In a further aspect, the present disclosure provides a method for inhibiting RNA polymerase in a subject in need thereof, comprising administering an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure to a subject in need thereof.

In a further aspect, the present disclosure provides use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure, in the manufacture of a medicament for treating a viral infection.

In a further aspect, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure, for treating a viral infection.

DETAILED DESCRIPTION OF THE DISCLOSURE

Reference will now be made in detail to certain embodiments of the present disclosure, examples of which are illustrated in the accompanying structures and formulas. While the present disclosure will be described in conjunction with the enumerated embodiments, it will be understood that they are not intended to limit the present disclosure to those embodiments. On the contrary, the present disclosure is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present disclosure as defined by the claims. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present disclosure. The present disclosure is in no way limited to the methods and materials described. In the event that one or more of the incorporated references and similar materials differs from or contradicts this application, including but not limited to defined terms, term usage, described techniques, or the like, the present disclosure controls. All references, patents, patent applications cited in the present disclosure are hereby incorporated by reference in their entireties.

It is appreciated that certain features of the present disclosure, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the present disclosure, which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable sub-combination. It must be noted that, as used in the specification and the appended claims, the singular forms “a, ” “an, ” and “the” include plural forms of the same unless the context clearly dictates otherwise. Thus, for example, reference to “acompound” includes a plurality of compounds.

Definitions

Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, 2^nd Edition, University Science Books, Sausalito, 2006; Smith and March March’s Advanced Organic Chemistry, 6^th Edition, John Wiley &Sons, Inc., New York, 2007; Larock, Comprehensive Organic Transformations, 3^rd Edition, VCH Publishers, Inc., New York, 2018; Carruthers, Some Modern Methods of Organic Synthesis, 4^th Edition, Cambridge University Press, Cambridge, 2004; the entire contents of each of which are incorporated herein by reference.

At various places in the present disclosure, linking substituents are described. It is specifically intended that each linking substituent includes both the forward and backward forms of the linking substituent. For example, -NR (CR’ R” ) -includes both -NR (CR’ R” ) -and - (CR’ R” ) NR-. Where the structure clearly requires a linking group, the Markush variables listed for that group are understood to be linking groups. For example, if the structure requires a linking group and the Markush group definition for that variable lists “alkyl” , then it is understood that the “alkyl” represents a linking alkylene group.

When a bond to a substituent is shown to cross a bond connecting two atoms in a ring, then such substituent may be bonded to any atom in the ring. When a substituent is listed without indicating the atom via which such substituent is bonded to the rest of the compound of a given formula, then such substituent may be bonded via any atom in such formula. Combinations of substituents and/or variables are permissible, but only if such combinations result in stable compounds.

When any variable (e.g., Rⁱ) occurs more than one time in any constituent or formula for a compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to be substituted with 0-2 Rⁱ moieties, then the group may optionally be substituted with up to two Rⁱ moieties and Rⁱ at each occurrence is selected independently from the definition of Rⁱ. Also, combinations of substituents and/or variables are permissible, but only if such combinations result in stable compounds.

As used herein, the term “C_i-j” indicates a range of the carbon atoms numbers, wherein i and j are integers and the range of the carbon atoms numbers includes the endpoints (i.e. i and j) and each integer point in between, and wherein j is greater than i. For examples, C_1-6 indicates a range of one to six carbon atoms, including one carbon atom, two carbon atoms, three carbon atoms, four carbon atoms, five carbon atoms and six carbon atoms. In some embodiments, the term “C_1-12” indicates 1 to 12, particularly 1 to 10, particularly 1 to 8, particularly 1 to 6, particularly 1 to 5, particularly 1 to 4, particularly 1 to 3 or particularly 1 to 2 carbon atoms.

As used herein, the term “alkyl” , whether as part of another term or used independently, refers to a saturated linear or branched-chain hydrocarbon radical, which may be optionally substituted independently with one or more substituents described herein. The term “C_i-j alkyl” refers to an alkyl having i to j carbon atoms. In some embodiments, alkyl groups contain 1 to 10 carbon atoms. In some embodiments, alkyl groups contain 1 to 9 carbon atoms. In some embodiments, alkyl groups contain 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of “C_1-10 alkyl” include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Examples of “C_1-6 alkyl” are methyl, ethyl, n-propyl, isopropyl, n-butyl, i-butyl, s-butyl, t-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2, 3-dimethyl-2-butyl, 3, 3-dimethyl-2-butyl, and the like. In some embodiments, alkyl groups contain 9 to 30 carbon atoms. In some embodiments, alkyl groups contain 9 to 28 carbon atoms, 9 to 26 carbon atoms, 9 to 24 carbon atoms, 10 to 28 carbon atoms, 10 to 26 carbon atoms, 10 to 24 carbon atoms, 12 to 28 carbon atoms, 12 to 26 carbon atoms, 12 to 24 carbon atoms, 14 to 28 carbon atoms, 14 to 26 carbon atoms, 14 to 24 carbon atoms, 14 to 22 carbon atoms, 14 to 20 carbon atoms, 14 to 18 carbon atoms, 14 to 16 carbon atoms, 16 to 22 carbon atoms, 16 to 20 carbon atoms, 16 to 18 carbon atoms, 18 to 22 carbon atoms, 18 to 20 carbon atoms, or 20 to 22 carbon atoms.

As used herein, the term “alkenyl” , whether as part of another term or used independently, refers to linear or branched-chain hydrocarbon radical having at least one carbon-carbon double bond, which may be optionally substituted independently with one or more substituents described herein, and includes radicals having “cis” and “trans” orientations, or alternatively, “E” and “Z” orientations. In some embodiments, alkenyl groups contain 2 to 12 carbon atoms. In some embodiments, alkenyl groups contain 2 to 11 carbon atoms. In some embodiments, alkenyl groups contain 2 to 11 carbon atoms, 2 to 10 carbon atoms, 2 to 9 carbon atoms, 2 to 8 carbon atoms, 2 to 7 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms. In some embodiments, alkenyl groups contain 9 to 30 carbon atoms. In some embodiments, alkenyl groups contain 9 to 28 carbon atoms, 9 to 26 carbon atoms, 9 to 24 carbon atoms, 10 to 28 carbon atoms, 10 to 26 carbon atoms, 10 to 24 carbon atoms, 12 to 28 carbon atoms, 12 to 26 carbon atoms, 12 to 24 carbon atoms, 14 to 28 carbon atoms, 14 to 26 carbon atoms, 14 to 24 carbon atoms, 14 to 22 carbon atoms, 14 to 20 carbon atoms, 14 to 18 carbon atoms, 14 to 16 carbon atoms, 16 to 22 carbon atoms, 16 to 20 carbon atoms, 16 to 18 carbon atoms, 18 to 22 carbon atoms, 18 to 20 carbon atoms, or 20 to 22 carbon atoms. Examples of alkenyl group include, but are not limited to, ethylenyl (or vinyl) , propenyl (allyl) , butenyl, pentenyl, 1-methyl-2 buten-1-yl, 5-hexenyl, and the like.

As used herein, the term “alkynyl” , whether as part of another term or used independently, refers to a linear or branched-chain hydrocarbon radical having at least one carbon-carbon triple bond, which may be optionally substituted independently with one or more substituents described herein. In some embodiments, alkenyl groups contain 2 to 12 carbon atoms. In some embodiments, alkynyl groups contain 2 to 11 carbon atoms. In some embodiments, alkynyl groups contain 2 to 11 carbon atoms, 2 to 10 carbon atoms, 2 to 9 carbon atoms, 2 to 8 carbon atoms, 2 to 7 carbon atoms, 2 to 6 carbon atoms, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms. In some embodiments, alkynyl groups contain 9 to 30 carbon atoms. In some embodiments, alkynyl groups contain 9 to 28 carbon atoms, 9 to 26 carbon atoms, 9 to 24 carbon atoms, 10 to 28 carbon atoms, 10 to 26 carbon atoms, 10 to 24 carbon atoms, 12 to 28 carbon atoms, 12 to 26 carbon atoms, 12 to 24 carbon atoms, 14 to 28 carbon atoms, 14 to 26 carbon atoms, 14 to 24 carbon atoms, 14 to 22 carbon atoms, 14 to 20 carbon atoms, 14 to 18 carbon atoms, 14 to 16 carbon atoms, 16 to 22 carbon atoms, 16 to 20 carbon atoms, 16 to 18 carbon atoms, 18 to 22 carbon atoms, 18 to 20 carbon atoms, or 20 to 22 carbon atoms. Examples of alkynyl group include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and the like.

As used herein, the term “amino” refers to -NH₂ group. Amino groups may also be substituted with one or more groups such as alkyl, aryl, carbonyl or other amino groups.

As used herein, the term “aryl” , whether as part of another term or used independently, refers to monocyclic and polycyclic ring systems having a total of 5 to 20 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains 3 to 12 ring members. Examples of “aryl” include, but are not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents. Also included within the scope of the term “aryl” , as it is used herein, is a group in which an aromatic ring is fused to one or more additional rings. In the case of polycyclic ring system, only one of the rings needs to be aromatic (e.g., 2, 3-dihydroindole) , although all of the rings may be aromatic (e.g., quinoline) . The second ring can also be fused or bridged. Examples of polycyclic aryl include, but are not limited to, benzofuranyl, indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like. Aryl groups can be substituted at one or more ring positions with substituents as described above.

As used herein, the term “azido” refers to -N₃.

As used herein, the term “cycloalkyl” , whether as part of another term or used independently, refer to a monovalent non-aromatic, saturated or partially unsaturated monocyclic and polycyclic ring system, in which all the ring atoms are carbon and which contains at least three ring forming carbon atoms. In some embodiments, the cycloalkyl may contain 3 to 12 ring forming carbon atoms, 3 to 10 ring forming carbon atoms, 3 to 9 ring forming carbon atoms, 3 to 8 ring forming carbon atoms, 3 to 7 ring forming carbon atoms, 3 to 6 ring forming carbon atoms, 3 to 5 ring forming carbon atoms, 4 to 12 ring forming carbon atoms, 4 to 10 ring forming carbon atoms, 4 to 9 ring forming carbon atoms, 4 to 8 ring forming carbon atoms, 4 to 7 ring forming carbon atoms, 4 to 6 ring forming carbon atoms, 4 to 5 ring forming carbon atoms. Cycloalkyl groups may be saturated or partially unsaturated. Cycloalkyl groups may be substituted. In some embodiments, the cycloalkyl group may be a saturated cyclic alkyl group. In some embodiments, the cycloalkyl group may be a partially unsaturated cyclic alkyl group that contains at least one double bond or triple bond in its ring system. In some embodiments, the cycloalkyl group may be monocyclic or polycyclic. Examples of monocyclic cycloalkyl group include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2- enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl. Examples of polycyclic cycloalkyl group include, but are not limited to, adamantyl, norbornyl, fluorenyl, spiro-pentadienyl, spiro [3.6] -decanyl, bicyclo [1, 1, 1] pentenyl, bicyclo [2, 2, 1] heptenyl, and the like.

As used herein, the term “cyano” refers to -CN.

As used herein, the term “halogen” refers to an atom selected from fluorine (or fluoro) , chlorine (or chloro) , bromine (or bromo) and iodine (or iodo) .

As used herein, the term “haloalkyl” refers to an alkyl substituted with one or more halogens. In some embodiments, the haloalkyl may contain 1 to 6 carbon atoms. In some embodiments, the haloalkyl may contain 1 to 4 carbon atoms. In some embodiments, the haloalkyl may contain 1 to 3 carbon atoms. Examples of haloalkyl include, but not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, chloromethyl, dichloromethyl, dibromomethyl, tribromomethyl and tetrafluoroethyl.

As used herein, the term “heteroatom” refers to nitrogen, oxygen, sulfur, phosphorus or silicon, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen (including N-oxides) .

As used herein, the term “heteroaryl” , whether as part of another term or used independently, refers to an aryl group having, in addition to carbon atoms, one or more heteroatoms. The heteroaryl group can be monocyclic. Examples of monocyclic heteroaryl include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, benzofuranyl and pteridinyl. The heteroaryl group also includes polycyclic groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Examples of polycyclic heteroaryl include, but are not limited to, indolyl, isoindolyl, benzothienyl, benzofuranyl, benzo [1, 3] dioxolyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

As used herein, the term “heterocyclyl” refers to a saturated or partially unsaturated carbocyclyl group in which one or more ring atoms are heteroatoms independently selected from oxygen, sulfur, nitrogen, phosphorus, silicon and the like, the remaining ring atoms being carbon, wherein one or more ring atoms may be optionally substituted independently with one or more substituents. In some embodiments, the heterocyclyl is a saturated heterocyclyl. In some embodiments, the heterocyclyl is a partially unsaturated heterocyclyl having one or more double bonds in its ring system. In some embodiments, the heterocyclyl may contains any oxidized form of carbon, nitrogen or sulfur, and any quaternized form of a basic nitrogen. “Heterocyclyl” also includes radicals wherein the heterocyclyl radicals are fused with a saturated, partially unsaturated, or fully unsaturated (i.e., aromatic) carbocyclic or heterocyclic ring. The heterocyclyl radical may be carbon linked or nitrogen linked where such is possible. In some embodiments, the heterocycle is carbon linked. In some embodiments, the heterocycle is nitrogen linked. For example, a group derived from pyrrole may be pyrrol-1-yl (nitrogen linked) or pyrrol-3-yl (carbon linked) . Further, a group derived from imidazole may be imidazol-1-yl (nitrogen linked) or imidazol-3-yl (carbon linked) .

In some embodiments, the term “3-to 12-membered heterocyclyl” refers to a 3-to 12-membered saturated or partially unsaturated monocyclic or polycyclic heterocyclic ring system having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. The fused, spiro and bridged ring systems are also included within the scope of this definition. Examples of monocyclic heterocyclyl include, but are not limited to oxetanyl, 1, 1-dioxothietanylpyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, thiazolyl, piperidyl, piperazinyl, piperidinyl, morpholinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, pyridonyl, pyrimidonyl, pyrazinonyl, pyrimidonyl, pyridazonyl, pyrrolidinyl, triazinonyl, and the like. Examples of fused heterocyclyl include, but are not limited to, phenyl fused ring or pyridinyl fused ring, such as quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, quinolizinyl, quinazolinyl, azaindolizinyl, pteridinyl, chromenyl, isochromenyl, indolyl, isoindolyl, indolizinyl, indazolyl, purinyl, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzothienyl, benzothiazolyl, carbazolyl, phenazinyl, phenothiazinyl, phenanthridinyl, imidazo [1, 2-a] pyridinyl, [1, 2, 4] triazolo [4, 3-a] pyridinyl, [1, 2, 3] triazolo [4, 3-a] pyridinyl groups, and the like. Examples of spiro heterocyclyl include, but are not limited to, spiropyranyl, spirooxazinyl, and the like. Examples of bridged heterocyclyl include, but are not limited to, morphanyl, hexamethylenetetraminyl, 3-aza-bicyclo [3.1.0] hexane, 8-aza-bicyclo [3.2.1] octane, 1-aza-bicyclo [2.2.2] octane, 1, 4-diazabicyclo [2.2.2] octane (DABCO) , and the like.

As used herein, the term “hydroxyl” refers to -OH.

As used herein, the term “metal ion” refers to a cation comprising a metal element. Various metal ions may be used as the metal ion in the present disclosure, including but not limited to any metal chosen from alkaline metals, alkaline earth metals, lanthanides, transition metals and mixtures thereof.

As used herein, the term “partially unsaturated” refers to a radical that includes at least one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aromatic (i.e., fully unsaturated) moieties.

As used herein, the term “protonated organic amine” refers to (R-NH₃) ⁺, wherein the R group refers to an organic radical.

As used herein, the term “purine base” or “pyrimidine base” comprises, but is not limited to, adenine, N⁶-alkylpurines, N⁶-acylpurines (wherein acyl is C (O) (alkyl, aryl, alkylaryl, or arylalkyl) ) , N⁶-benzylpurine, N⁶-halopurine, N⁶-vinylpurine, N⁶-acetylenic purine, N⁶-acyl purine, N⁶-hydroxyalkyl purine, N⁶-allylaminopurine, N⁶-thioallyl purine, N²-alkylpurines, N²-alkyl-6-thiopurines, thymine, cytosine, 5-fluorocytosine, 5-methylcytosine, 6-azapyrimidine, including 6-azacytosine, 2-and/or 4-mercaptopyrmidine, uracil, 5-halouracil, including 5-fluorouracil, C⁵-alkylpyrimidines, C⁵-benzylpyrimidines, C⁵-halopyrimidines, C⁵-vinylpyrimidine, C⁵-acetylenic pyrimidine, C⁵-acyl pyrimidine, C⁵-hydroxyalkyl purine, C⁵-amidopyrimidine, C⁵-cyanopyrimidine, C⁵-5-iodopyrimidine, C⁶-iodo-pyrimidine, C⁵-Br-vinyl pyrimidine, C⁶-Br-vinyl pyriniidine, C⁵-nitropyrimidine, C⁵-amino-pyrimidine, N²-alkylpurines, N²-alkyl-6-thiopurines, 5-azacytidinyl, 5-azauracilyl, triazolopyridinyl, imidazolopyridinyl, pyrrolopyrimidinyl, and pyrazolopyrimidinyl. Purine bases include, but are not limited to, guanine, adenine, hypoxanthine, 2, 6-diaminopurine, and 6-chloropurine. The purine and pyrimidine bases are linked to the ribose sugar, or analog thereof, through a nitrogen atom of the base. Functional oxygen and nitrogen groups on the base can be protected as necessary or desired. Suitable protecting groups are well known to those skilled in the art, and include trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, and t-butyldiphenylsilyl, trityl, alkyl groups, and acyl groups such as acetyl and propionyl, methanesulfonyl, and p-toluenesulfonyl.

As used herein, the term “substituted” , whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and that the substitution results in a stable or chemically feasible compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as “unsubstituted” , references to chemical moieties herein are understood to include substituted variants. For example, reference to an “aryl” group or moiety implicitly includes both substituted and unsubstituted variants.

Compounds

The present disclosure provides novel compounds of Formula (I) and pharmaceutically acceptable salts thereof, synthetic methods for making the compounds, pharmaceutical compositions containing them and various uses of the disclosed compounds.

In one aspect, the present disclosure provides a compound having Formula (I) :

or a pharmaceutically acceptable salt thereof,

wherein

Base is a naturally occurring or modified pyrimidine base or purine base;

Q is CH₂, CHD or CD₂;

L is selected from hydrogen or -C (R⁵) ₂-L¹-L²;

L² is selected from hydrogen, R⁶, wherein *end of L² is connected to L¹;

each R^a is independently selected from hydrogen, halogen or alkyl;

m is 0, 1, 2, or 3; and

each n, p, or q is independently an integer between 0-20.

In another aspect, there is provided a compound having a Formula (Ia) :

or a pharmaceutically acceptable salt thereof, wherein L, R¹-R⁴, M, base and m are defined supra.

In some embodiments, the base is selected from the group consisting of:

In certain embodiments, the base is selected from the group consisting of:

In some embodiments, L is hydrogen.

In certain embodiments, m is 0, 1 or 3.

In certain embodiments, M is hydrogen.

In some embodiments, L is-C (R⁵) ₂-L¹-L².

In some embodiments, both R⁵ are hydrogen or deuterium.

In certain embodiments, one R⁵ is hydrogen, and the other R⁵ is alkyl optionally substituted with aryl.

In certain embodiments, one R⁵ is hydrogen, and the other R⁵ is methyl, ethyl,

In some embodiments, L¹ is selected from -OC (O) -*, -OC (O) O-*, -OC (O) NH-*, -OC (O) N (CH₃) -*, -NHC (O) -*, -N (CH₃) C (O) -*, -NHC (O) O-*, -N (CH₃) C (O) O-*, or

In certain embodiments, L¹ is -OC (O) -*, -OC (O) O-*or

In some embodiments, L² is R⁶.

In some embodiments, R⁶ is linear C_9-26 alkyl, linear C_9-26 alkenyl, linear C_9- ₂₆ alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-.

In certain embodiments, R⁶ is linear C_14-22 alkyl. In certain embodiments, R⁶ is linear C_16-20 alkyl.

In certain embodiments, R⁶ is linear C_14-22 alkenyl. In certain embodiments, R⁶ is linear C_16-20 alkenyl.

In certain embodiments, R⁶ is linear C_14-22 alkenyl comprising 1-6 double bonds.

In certain embodiments, R⁶ is linear C_14-22 alkenyl comprising 1-6 double bonds and each double bond is in Z configuration.

In certain embodiments, R⁶ is selected from the group consisting of:

In some embodiments, R⁶ is CH₃ (CH₂) _pO (CH₂) _q-.

In certain embodiments, R⁶ is CH₃ (CH₂) _pO (CH₂) _q-, wherein p is an integer from 3 to 20, and q is an integer from 1 to 6.

In certain embodiments, R⁶ is selected from CH₃ (CH₂) ₅O (CH₂) ₂-, CH₃ (CH₂) ₅O (CH₂) ₃-, CH₃ (CH₂) ₅O (CH₂) ₄-, CH₃ (CH₂) ₅O (CH₂) ₅-, CH₃ (CH₂) ₆O (CH₂) ₃-, CH₃ (CH₂) ₇O (CH₂) ₃-, CH₃ (CH₂) ₈O (CH₂) ₃-, CH₃ (CH₂) ₉O (CH₂) ₃-, CH₃ (CH₂) ₁₀O (CH₂) ₃-, CH₃ (CH₂) ₁₁O (CH₂) ₃-, CH₃ (CH₂) ₁₂O (CH₂) ₃-, CH₃ (CH₂) ₁₃O (CH₂) ₃-, CH₃ (CH₂) ₁₄O (CH₂) ₃-, CH₃ (CH₂) ₁₅O (CH₂) ₃-, CH₃ (CH₂) ₁₆O (CH₂) ₃-, CH₃ (CH₂) ₁₇O (CH₂) ₃-, CH₃ (CH₂) ₁₈O (CH₂) ₃-, or CH₃ (CH₂) ₁₉O (CH₂) ₃-.

In some embodiments, R⁶ is R^aOCH₂ (CH₂OCH₂) _nCH₂-.

In certain embodiments, R⁶ is R^aOCH₂ (CH₂OCH₂) _nCH₂-and n is an integer from 1 to 6.

In certain embodiments, R⁶ is R^aOCH₂ (CH₂OCH₂) _nCH₂-, wherein R^a is hydrogen, methyl or ethyl.

In certain embodiments, R⁶ is selected from the group consisting of CH₃OCH₂ (CH₂OCH₂) ₂CH₂-, CH₃OCH₂ (CH₂OCH₂) ₃CH₂-, CH₃OCH₂ (CH₂OCH₂) ₄CH₂-, HOCH₂ (CH₂OCH₂) ₅CH₂-, CH₃OCH₂ (CH₂OCH₂) ₅CH₂-, and CH₃CH₂OCH₂ (CH₂OCH₂) ₅CH₂-.

In some embodiments, L² is

In some embodiments, R⁷ is C_6-26 alkyl, C_6-26 alkenyl, C_6-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-.

In certain embodiments, R⁷ is linear C_6-26 alkyl or linear C_6-26 alkenyl.

In certain embodiments, R⁷ is linear C_13-21 alkyl or linear C_13-21 alkenyl. In certain embodiments, R⁷ is linear C_15-20 alkyl or linear C_15-20 alkenyl.

In certain embodiments, R⁷ is linear C_13-21 alkenyl comprising 1-6 double bonds.

In certain embodiments, R⁷ is linear C_13-21 alkenyl comprising 1-6 double bonds and each double bond is in Z configuration.

In certain embodiments, R⁷ is selected from the group consisting of:

In some embodiments, R⁷ is CH₃ (CH₂) _pO (CH₂) _q-.

In certain embodiments, R⁷ is CH₃ (CH₂) _pO (CH₂) _q-, wherein p is an integer from 3 to 20, and q is an integer from 1 to 6.

In certain embodiments, R⁷ is selected from CH₃ (CH₂) ₅O (CH₂) ₂-, CH₃ (CH₂) ₅O (CH₂) ₃-, CH₃ (CH₂) ₅O (CH₂) ₄-, CH₃ (CH₂) ₅O (CH₂) ₅-, CH₃ (CH₂) ₆O (CH₂) ₃-, CH₃ (CH₂) ₇O (CH₂) ₃-, CH₃ (CH₂) ₈O (CH₂) ₃-, CH₃ (CH₂) ₉O (CH₂) ₃-, CH₃ (CH₂) ₁₀O (CH₂) ₃-, CH₃ (CH₂) ₁₁O (CH₂) ₃-, CH₃ (CH₂) ₁₂O (CH₂) ₃-, CH₃ (CH₂) ₁₃O (CH₂) ₃-, CH₃ (CH₂) ₁₄O (CH₂) ₃-, CH₃ (CH₂) ₁₅O (CH₂) ₃-, CH₃ (CH₂) ₁₆O (CH₂) ₃-, CH₃ (CH₂) ₁₇O (CH₂) ₃-, CH₃ (CH₂) ₁₈O (CH₂) ₃-, or CH₃ (CH₂) ₁₉O (CH₂) ₃-.

In some embodiments, R⁷ is R^aOCH₂ (CH₂OCH₂) _nCH₂-.

In certain embodiments, R⁷ is R^aOCH₂ (CH₂OCH₂) _nCH₂-, wherein n is an integer from 1 to 6.

In certain embodiments, R⁷ is R^aOCH₂ (CH₂OCH₂) _nCH₂-, wherein R^a is hydrogen, methyl or ethyl.

In certain embodiments, R⁷ is selected from the group consisting of CH₃OCH₂ (CH₂OCH₂) ₂CH₂-, CH₃OCH₂ (CH₂OCH₂) ₃CH₂-, CH₃OCH₂ (CH₂OCH₂) ₄CH₂-, HOCH₂ (CH₂OCH₂) ₅CH₂-, CH₃OCH₂ (CH₂OCH₂) ₅CH₂-, and CH₃CH₂OCH₂ (CH₂OCH₂) ₅CH₂-.

In some embodiments, R⁸ is C_1-26 alkyl or C_2-26 alkenyl, and R⁹ is hydrogen or C_1-26 alkyl.

In certain embodiments, R⁸ is linear C_1-26 alkyl or linear C_8-26 alkenyl, and R⁹ is hydrogen or linear C_1-26 alkyl.

In certain embodiments, R⁷ is linear C_6-26 alkyl, R⁸ is linear C_2-26 alkyl, and R⁹ is hydrogen.

In certain embodiments, R⁷ is linear C_7-26 alkyl, R⁸ is methyl, and R⁹ is hydrogen or methyl.

In some embodiments, L² is

In certain embodiments, R⁷ is linear C_6-26 alkyl, linear C_6-26 alkenyl, linear C_6- ₂₆ alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-.

In certain embodiments, R⁷ is linear C_6-26 alkyl or linear C_6-26 alkenyl.

In certain embodiments, R⁸ is linear C_1-26 alkyl, linear C_2-26 alkenyl, linear C_2- ₂₆ alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-, and R⁹ is hydrogen or C_1-26 alkyl.

In certain embodiments, R⁸ is linear C_1-26 alkyl or linear C_2-26 alkenyl, and R⁹ is hydrogen or linear C_1-26 alkyl.

In certain embodiments, R⁷ is linear C_6-26 alkyl, R⁸ is linear C_1-26 alkyl, and R⁹ is hydrogen.

In certain embodiments, R⁷ and R⁸ are delineated as follows:

In some embodiments, R¹ is selected from hydrogen, hydroxyl, cyano, azido, alkyl, or haloalkyl.

In certain embodiments, R¹ is cyano.

In certain embodiments, R¹ is alkyl. In certain embodiments, R¹ is methyl.

In certain embodiments, R¹ is haloalkyl. In certain embodiments, R¹ is -CH₂F.

In some embodiments, each of R²¹, R²², R³¹, R³², and R⁴ is independently selected from hydrogen, deuterium, halogen, hydroxyl, cyano, azido, alkyl, haloalkyl, -OC (O) R^a.

In certain embodiments, each of R²¹, R²², R³¹, R³² is selected from hydrogen, deuterium, halogen, hydroxyl, alkyl or -OC (O) R^a. In certain embodiments, R^a is alkyl. In certain embodiments, R^a is methyl.

In certain embodiments, R⁴ is selected from hydrogen, deuterium, halogen, cyano, azido, or haloalkyl.

In some embodiments, is selected from the group consisting of:

In some embodiments, the base isand is

In certain embodiments, the base is

In a further aspect, the present disclosure provides a compound having a formula selected from the group consisting of:

Compounds provided herein are described with reference to both generic formulae and specific compounds. In addition, compounds of the present disclosure may exist in a number of different forms or derivatives, all within the scope of the present disclosure. These include, for example, tautomers, stereoisomers, racemic mixtures, regioisomers, salts, solvated forms, amorphous forms, different crystal forms or polymorphs.

The compounds of present disclosure can comprise one or more asymmetric centers depending on substituent selection, and thus can exist in various stereoisomeric forms, e.g., enantiomers and/or diastereomers. For example, the compounds provided herein may have an asymmetric carbon center, and thus compounds provided herein may have either the (R) or (S) stereo-configuration at a carbon asymmetric center. Therefore, compounds of the present disclosure may be in the form of an individual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers.

As used herein, the term “enantiomer” refers to two stereoisomers of a compound which are non-superimposable mirror images of one another. The term “diastereomer” refers to a pair of optical isomers which are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.

Where a particular enantiomer is preferred, it may, in some embodiments be provided substantially free of the opposite enantiomer, and may also be referred to as “optically enriched” . “Optically enriched” , as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments, the compound is made up of at least about 90%by weight of a preferred enantiomer. In other embodiments, the compound is made up of at least about 95%, 98%, or 99%by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, for example by chromatography or crystallization, by the use of stereochemically uniform starting materials for the synthesis or by stereoselective synthesis. Optionally a derivatization can be carried out before a separation of stereoisomers. The separation of a mixture of stereoisomers can be carried out at an intermediate step during the synthesis of a compound provided herein or it can be done on a final racemic product. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing a stereogenic center of known configuration. Alternatively, absolute stereochemistry may be determined by Vibrational Circular Dichroism (VCD) spectroscopy analysis. See, for example, Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981) ; Wilen, S.H., et al., Tetrahedron 33: 2725 (1977) ; Eliel, E.L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962) ; Wilen, S.H. Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972) .

In some embodiments, mixtures of diastereomers, for example mixtures of diastereomers enriched with 51%or more of one of the diastereomers, including for example 60%or more, 70%or more, 80%or more, or 90%or more of one of the diastereomers are provided.

In some embodiments, compounds provided herein may have one or more double bonds that can exist as either the Z or E isomer, unless otherwise indicated. The present disclosure additionally encompasses the compounds as individual isomers substantially free of other isomers and alternatively, as mixtures of various isomers, e.g., racemic mixtures of enantiomers.

The compounds of the present disclosure may also exist in different tautomeric forms, and all such forms are embraced within the scope of the present disclosure. The term “tautomer” or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol, amide-imidic acid, lactam-lactim, imine-enamine isomerizations and annular forms where a proton can occupy two or more positions of a heterocyclic system (for example, 1H-and 3H-imidazole, 1H-, 2H-and 4H-1, 2, 4-triazole, 1H-and 2H-isoindole, and 1H-and 2H-pyrazole) . Valence tautomers include interconversions by reorganization of some of the bonding electrons. Tautomers can be in equilibrium or sterically locked into one form by appropriate substitution. Compounds of the present disclosure identified by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

The present disclosure is also intended to include all isotopes of atoms in the compounds. Isotopes of an atom include atoms having the same atomic number but different mass numbers. For example, unless otherwise specified, hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, bromide or iodine in the compounds of present disclosure are meant to also include their isotopes, such as but not limited to ¹H, ²H, ³H, ¹¹C, ¹²C, ¹³C, ¹⁴C, ¹⁴N, ¹⁵N, ¹⁶O, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³²S, ³³S, ³⁴S, ³⁶S, ¹⁷F, ¹⁸F, ¹⁹F, ³⁵Cl, ³⁷Cl, ⁷⁹Br, ⁸¹Br, ¹²⁴I, ¹²⁷I and ¹³¹I. Isotopically-enriched compounds of Formula (I) can be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the Schemes and Examples herein using appropriate isotopically-enriched reagents and/or intermediates.

In some embodiments, the present disclosure includes compounds of Formula (I) or (Ia) wherein one or more hydrogens attached to a carbon atom is/are replaced by deuterium. Such compounds exhibit increased resistance to metabolism and are thus useful for increasing the half-life of any compound of Formula (I) or (Ia) when administered to a subject, such as mammal, particularly a human. See, for example, Foster, “Deuterium Isotope Effects in Studies of Drug Metabolism” , Trends Pharmacol. Sci. 5 (12) : 524-527 (1984) . In view of the present disclosure, such compounds are synthesized by means known in the art, for example by employing starting materials in which one or more hydrogens have been replaced by deuterium.

Also falling within the scope herein are the in vivo metabolic products of the compounds described herein, to the extent such products are novel and unobvious over the prior art. Such products may result for example from the oxidation, reduction, hydrolysis, amidation, esterification and the like of the administered compound, primarily due to enzymatic processes. Accordingly, included are novel and unobvious compounds produced by a process comprising contacting a compound with a mammal for a period of time sufficient to yield a metabolic product thereof.

Compounds of the present disclosure can be formulated as or be in the form of pharmaceutically acceptable salts. Unless specified to the contrary, a compound provided herein includes pharmaceutically acceptable salts of such compound.

As used herein, the term “pharmaceutically acceptable” indicates that the substance or composition is compatible chemically and/or toxicologically, with the other ingredients comprising a formulation, and/or the subjects being treated therewith.

As used herein, the term “pharmaceutically acceptable salt” , unless otherwise indicated, includes salts that retain the biological effectiveness of the free acids and bases of the specified compound and that are not biologically or otherwise undesirable. Contemplated pharmaceutically acceptable salt forms include, but are not limited to, mono, bis, tris, tetrakis, and so on. Pharmaceutically acceptable salts are non-toxic in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing the solubility to facilitate administering higher concentrations of the drug.

Pharmaceutically acceptable salts include acid addition salts such as those containing sulfate, chloride, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate and quinate. Pharmaceutically acceptable salts can be obtained from acids such as hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

Pharmaceutically acceptable salts also include basic addition salts such as those containing benzathine, chloroprocaine, choline, diethanolamine, ethanolamine, t-butylamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc, when acidic functional groups, such as carboxylic acid or phenol are present. For example, see Remington's Pharmaceutical Sciences, 19^thed., Mack Publishing Co., Easton, PA, Vol. 2, p. 1457, 1995; “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth, Wiley-VCH, Weinheim, Germany, 2002. Such salts can be prepared using the appropriate corresponding bases.

Pharmaceutically acceptable salts can be prepared by standard techniques. For example, the free-base form of a compound can be dissolved in a suitable solvent, such as an aqueous or aqueous-alcohol solution containing the appropriate acid and then isolated by evaporating the solution. Thus, if the particular compound is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.

Similarly, if the particular compound is an acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary) , an alkali metal hydroxide or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include organic salts derived from amino acids, such as L-glycine, L-lysine, and L-arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as hydroxyethylpyrrolidine, piperidine, morpholine or piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

It is also to be understood that the compounds of present disclosure can exist in unsolvated forms, solvated forms (e.g., hydrated forms) , and solid forms (e.g., crystal or polymorphic forms) , and the present disclosure is intended to encompass all such forms.

As used herein, the term “solvate” or “solvated form” refers to solvent addition forms that contain either stoichiometric or non-stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate; and if the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of the substance in which the water retains its molecular state as H₂O. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

As used herein, the terms “crystal form” , “crystalline form” , “polymorphic forms” and “polymorphs” can be used interchangeably, and mean crystal structures in which a compound (or a salt or solvate thereof) can crystallize in different crystal packing arrangements, all of which have the same elemental composition. Different crystal forms usually have different X-ray diffraction patterns, infrared spectral, melting points, density hardness, crystal shape, optical and electrical properties, stability and solubility. Recrystallization solvent, rate of crystallization, storage temperature, and other factors may cause one crystal form to dominate. Crystal polymorphs of the compounds can be prepared by crystallization under different conditions.

Synthesis of compounds

Synthesis of the compounds provided herein, including pharmaceutically acceptable salts thereof, are illustrated in the synthetic schemes in the examples. The compounds provided herein can be prepared using any known organic synthesis techniques and can be synthesized according to any of numerous possible synthetic routes, and thus these schemes are illustrative only and are not meant to limit other possible methods that can be used to prepare the compounds provided herein. Additionally, the steps in the Schemes are for better illustration and can be changed as appropriate. The embodiments of the compounds in examples were synthesized for the purposes of research and potentially submission to regulatory agencies.

The reactions for preparing compounds of the present disclosure can be carried out in suitable solvents, which can be readily selected by one skilled in the art of organic synthesis. Suitable solvents can be substantially non-reactive with the starting materials (reactants) , the intermediates, or products at the temperatures at which the reactions are carried out, e.g. temperatures that can range from the solvent’s freezing temperature to the solvent's boiling temperature. A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction step, suitable solvents for a particular reaction step can be selected by one skilled in the art.

Preparation of compounds of the present disclosure can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups, can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., Wiley &Sons, Inc., New York (1999) , in P. Kocienski, Protecting Groups, Georg Thieme Verlag, 2003, and in Peter G.M. Wuts, Greene's Protective Groups in Organic Synthesis, 5^th Edition, Wiley, 2014, all of which are incorporated herein by reference in its entirety.

Reactions can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g. ¹H or ¹³C) , infrared spectroscopy, spectrophotometry (e.g. UV-visible) , mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC) , liquid chromatography-mass spectroscopy (LCMS) , or thin layer chromatography (TLC) . Compounds can be purified by one skilled in the art by a variety of methods, including high performance liquid chromatography (HPLC) ( “Preparative LC-MS Purification: Improved Compound Specific Method Optimization” Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6 (6) , 874-883, which is incorporated herein by reference in its entirety) , and normal phase silica chromatography.

The known starting materials of the present disclosure can be synthesized by using or according to the known methods in the art, or can be purchased from commercial suppliers. Unless otherwise noted, analytical grade solvents and commercially available reagents were used without further purification.

Unless otherwise specified, the reactions of the present disclosure were all done under a positive pressure of nitrogen or argon or with a drying tube in anhydrous solvents, and the reaction flasks were typically fitted with rubber septa for the introduction of substrates and reagents via syringe. Glassware was oven dried and/or heat dried.

For illustrative purposes, the Examples section below shows synthetic route for preparing the compounds of the present disclosure as well as key intermediates. Those skilled in the art will appreciate that other synthetic routes may be used to synthesize the inventive compounds. Although specific starting materials and reagents are depicted, other starting materials and reagents can be easily substituted to provide a variety of derivatives and/or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified in light of this disclosure using conventional chemistry well known to those skilled in the art.

Use of Compounds

In an aspect, the present disclosure provides compounds of Formula (I) or pharmaceutically acceptable salts thereof, which are capable of inhibiting RNA polymerase. Thus, the compounds of the present disclosure or a pharmaceutically acceptable salt thereof are useful as medicinal drugs, and particularly useful as therapeutic or prophylactic agent that are active against various viruses. In some embodiments, the compounds of the present disclosure are therapeutic or prophylactic agent active against caliciviruses, picornaviruses and coronaviruses.

As used herein, the term “therapy” is intended to have its normal meaning of dealing with a disease in order to entirely or partially relieve one, some or all of its symptoms, or to correct or compensate for the underlying pathology, thereby achieving beneficial or desired clinical results. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total) , whether detectable or undetectable. “Therapy” can also mean prolonging survival as compared to expected survival if not receiving it. Those in need of therapy include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented. The term “therapy” also encompasses prophylaxis unless there are specific indications to the contrary. The terms “therapeutic” and “therapeutically” should be interpreted in a corresponding manner.

The term “treatment” is used synonymously with “therapy” . Similarly the term “treat” can be regarded as “applying therapy” where “therapy” is as defined herein.

As used herein, the term “prophylaxis” is intended to have its normal meaning and includes primary prophylaxis to prevent the development of the disease and secondary prophylaxis whereby the disease has already developed and the patient is temporarily or permanently protected against exacerbation or worsening of the disease or the development of new symptoms associated with the disease.

In a further aspect, the present disclosure provides use of the compound of the present disclosure or a pharmaceutically acceptable salt thereof for treatment of viral infection.

In a further aspect, the present disclosure provides use of the compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure, in the manufacture of a medicament for treating a viral infection.

In some embodiments, the compounds of the present disclosure have good inhibitory effect against various coronaviruses. In some embodiments, the compounds of the present disclosure have good inhibitory effect against various coronaviruses with low cytotoxicity.

In some embodiments, the compounds of the present disclosure can convert into triphosphate and maintain a high concentration of triphosphate in targeted tissue and celles, such as lung cell and respiratory epidermal cell.

Therefore, the present disclosure provides use of the compound of the present disclosure or a pharmaceutically acceptable salt thereof for treatment of coronavirus infection.

In a further aspect, the present disclosure provides use of the compound of the present disclosure or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of the present disclosure, in the manufacture of a medicament for treating a coronavirus infection.

Pharmaceutical Compositions

For the purposes of administration, in some embodiments, the compounds provided herein are administered as a raw chemical or are formulated as pharmaceutical compositions.

Therefore, in a further aspect, there is provided pharmaceutical compositions comprising one or more compounds of the present disclosure, or a pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutical compositions of the present disclosure comprise a compound selected from any one of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical compositions of the present disclosure comprise a first compound selected from any one of Formula (I) or a pharmaceutically acceptable salt thereof and one or more additional compounds of the same formula but said first compound and additional compounds are not the same molecules.

As used herein, the term “pharmaceutical composition” refers to a formulation containing the molecules or compounds of the present disclosure in a form suitable for administration to a subject.

In some embodiments, the pharmaceutical compositions of the present disclosure comprises a therapeutically effective amount of one or more compounds of Formula (I) or a pharmaceutically acceptable salt thereof.

As used herein, the term “therapeutically effective amount” refers to an amount of a molecule, compound, or composition comprising the molecule or compound to treat, ameliorate, or prevent an identified disease or condition, or to exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend upon the subject’s body weight, size, and health; the nature and extent of the condition; the rate of administration; the therapeutic or combination of therapeutics selected for administration; and the discretion of the prescribing physician. Therapeutically effective amounts for a given situation can be determined by routine experimentation that is within the skill and judgment of the clinician.

In another aspect, there is provided pharmaceutical composition comprising one or more compounds of the present disclosure, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical acceptable excipient.

As used herein, the term “pharmaceutically acceptable excipient” refers to an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes excipient that is acceptable for veterinary use as well as human pharmaceutical use. A “pharmaceutically acceptable excipient” as used herein includes both one and more than one such excipient. The term “pharmaceutically acceptable excipient” also encompasses “pharmaceutically acceptable carrier” and “pharmaceutically acceptable diluent” .

The particular excipient used will depend upon the means and purpose for which the compounds of the present disclosure is being applied. Solvents are generally selected based on solvents recognized by persons skilled in the art as safe to be administered to a mammal including humans. In general, safe solvents are non-toxic aqueous solvents such as water and other non-toxic solvents that are soluble or miscible in water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycols (e.g., PEG 400, PEG 300) , etc. and mixtures thereof.

In some embodiments, suitable excipients may include buffers such as phosphate, citrate and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol) ; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose, dextrins, or substituted dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes) ; and/or non-ionic surfactants such as TWEEN^TM, PLURONICS^TM or polyethylene glycol (PEG) .

In some embodiments, suitable excipients may include one or more stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present disclosure or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament) . The active pharmaceutical ingredients may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly- (methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980) . A “liposome” is a small vesicle composed of various types of lipids, phospholipids and/or surfactant which is useful for delivery of a drug (such as the compounds disclosed herein and, optionally, a chemotherapeutic agent) to a mammal including humans. The components of the liposome are commonly arranged in a bilayer formation, similar to the lipid arrangement of biological membranes.

The pharmaceutical compositions provided herein can be in any form that allows for the composition to be administered to a subject, including, but not limited to a human, and formulated to be compatible with an intended route of administration.

A variety of routes are contemplated for the pharmaceutical compositions provided herein, and accordingly the pharmaceutical composition provided herein may be supplied in bulk or in unit dosage form depending on the intended administration route. For example, for oral, buccal, and sublingual administration, powders, suspensions, granules, tablets (such as orodispersible tablets or orally disintegrating tablets) , pills, oral soluble films, capsules, gelcaps, and caplets may be acceptable as solid dosage forms, and emulsions, syrups, elixirs, suspensions, and solutions may be acceptable as liquid dosage forms. For injection administration, emulsions and suspensions may be acceptable as liquid dosage forms, and a powder suitable for reconstitution with an appropriate solution as solid dosage forms. For inhalation administration, solutions, sprays, dry powders, and aerosols may be acceptable dosage form. For topical (including buccal and sublingual) or transdermal administration, powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches may be acceptable dosage form. For vaginal administration, pessaries, tampons, creams, gels, pastes, foams and spray may be acceptable dosage form. For nasal or pulmonary administration, aqueous or oily solutions may be acceptable dosage form.

The quantity of active ingredient in a unit dosage form of composition is a therapeutically effective amount and is varied according to the particular treatment involved. As used herein, the term “therapeutically effective amount” refers to an amount of a molecule, compound, or composition comprising the molecule or compound to treat, ameliorate, or prevent an identified disease or condition, or to exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The precise effective amount for a subject will depend upon the subject’s body weight, size, and health; the nature and extent of the condition; the rate of administration; the therapeutic or combination of therapeutics selected for administration; and the discretion of the prescribing physician. Therapeutically effective amounts for a given situation can be determined by routine experimentation that is within the skill and judgment of the clinician.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for oral administration.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of tablet formulations. Suitable pharmaceutically-acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or algenic acid; binding agents such as starch; lubricating agents such as magnesium stearate, stearic acid or talc; preservative agents such as ethyl or propyl p-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case using conventional coating agents and procedures well known in the art.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in a form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of aqueous suspensions, which generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate) , or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid) , coloring agents, flavoring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame) .

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of oily suspensions, which generally contain suspended active ingredient in a vegetable oil (such as arachis oil, castor oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin) . The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these. Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavoring and preservative agents.

In certain embodiments, the pharmaceutical compositions provided herein may be in the form of syrups and elixirs, which may contain sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, a demulcent, a preservative, a flavoring and/or coloring agent.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for injection administration.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents, which have been mentioned above. The sterile injectable preparation may 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 vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for nasal or pulmonary administration. In certain embodiments, the formulation is administered by rapid inhalation through the nasal passage or by inhalation through the mouth so as to reach the alveolar sacs.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for inhalation administration.

In certain embodiments, the pharmaceutical compositions of the present disclosure may be in the form of aqueous and nonaqueous (e.g., in a fluorocarbon propellant) aerosols containing any appropriate solvents and optionally other compounds such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH modifiers, surfactants, bioavailability modifiers and combinations of these. The carriers and stabilizers vary with the requirements of the particular compound, but typically include nonionic surfactants (Tweens, Pluronics, or polyethylene glycol) , innocuous proteins like serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars or sugar alcohols.

In some embodiments, the pharmaceutical compositions of the present disclosure may be in a form of formulation for topical or transdermal administration.

In certain embodiments, the pharmaceutical compositions provided herein may be in the form of creams, ointments, gels and aqueous or oily solutions or suspensions, which may generally be obtained by formulating an active ingredient with a conventional, topically acceptable excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

In certain embodiments, the pharmaceutical compositions provided herein may be formulated in the form of transdermal skin patches that are well known to those of ordinary skill in the art.

Besides those representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the present disclosure. Such excipients and carriers are described, for example, in “Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991) , in “Remington: The Science and Practice of Pharmacy” , Ed. University of the Sciences in Philadelphia, 21^st Edition, LWW (2005) , which are incorporated herein by reference.

In some embodiments, the pharmaceutical compositions of the present disclosure can be formulated as a unit dosage form. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. The amount of the compounds provided herein in the unit dosage form will vary depending on the condition to be treated, the subject to be treated (e.g., the age, weight, and response of the individual subject) , the particular route of administration, the actual compound administered and its relative activity, and the severity of the subject's symptoms.

In some embodiments, dosage levels of the pharmaceutical compositions of the present disclosure can be between 0.001-1000 mg/kg body weight/day, for example, 0.001-1000 mg/kg body weight/day, 0.001-900 mg/kg body weight/day, 0.001-800 mg/kg body weight/day, 0.001-700 mg/kg body weight/day, 0.001-600 mg/kg body weight/day, 0.001-500 mg/kg body weight/day, 0.001-400 mg/kg body weight/day, 0.001-300 mg/kg body weight/day, 0.001-200 mg/kg body weight/day, 0.001-100 mg/kg body weight/day, 0.001-50 mg/kg body weight/day, 0.001-40 mg/kg body weight/day, 0.001-30 mg/kg body weight/day, 0.001-20 mg/kg body weight/day, 0.001-10 mg/kg body weight/day, 0.001-5 mg/kg body weight/day, 0.001-1 mg/kg body weight/day, 0.001-0.5 mg/kg body weight/day, 0.001-0.4 mg/kg body weight/day, 0.001-0.3 mg/kg body weight/day, 0.001-0.2 mg/kg body weight/day, 0.001-0.1 mg/kg body weight/day, 0.005-0.1 mg/kg body weight/day, 0.01-0.1 mg/kg body weight/day, 0.02-0.1 mg/kg body weight/day, 0.03-0.1 mg/kg body weight/day, 0.04-0.1 mg/kg body weight/day, 0.05-0.1 mg/kg body weight/day, 0.06-0.1 mg/kg body weight/day, 0.07-0.1 mg/kg body weight/day, 0.08-0.1 mg/kg body weight/day, or 0.09-0.1 mg/kg body weight/day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, provided that such larger doses are first divided into several small doses for administration throughout the day. For further information on routes of administration and dosage regimes, see Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board) , Pergamon Press 1990, which is specifically incorporated herein by reference.

In some embodiments, the pharmaceutical compositions of the present disclosure are formulated for oral administration. In some embodiments, the unit dosage for oral administration contains one or more compounds provided herein in an amount from about 1 mg to about 1000 mg, for example from about 5 mg to about 1000 mg, from about 10 mg to about 1000 mg, from about 15 mg to about 1000 mg, from about 20 mg to about 1000 mg, from about 25 mg to about 1000 mg, from about 30 mg to about 1000 mg, from about 40 mg to about 1000 mg, from about 50 mg to about 1000 mg, from about 60 mg to about 1000 mg, from about 70 mg to about 1000 mg, from about 80 mg to about 1000 mg, from about 90 mg to about 1000 mg, from about 100 mg to about 1000 mg, from about 200 mg to 1000 mg, from about 300 mg to about 1000 mg, from about 400 mg to about 1000 mg, from about 500 mg to about 1000 mg, from about 1 mg to 500 mg, from about 10 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 200 mg to about 500 mg, from about 300 mg to about 500 mg, from about 400 mg to about 500 mg, for example about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg and the like. In some embodiments, the dosage unit may be administered to a subject from 1 to 6 times per day depending on the severity of the subject’s symptoms.

In some embodiments, the pharmaceutical compositions of the present disclosure are formulated for oral administration in a treatment having a duration of more than 1 week, more than 2 weeks, more than 3 weeks, more than 1 month, more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 7 months, more than 8 months, more than 9 months, more than 10 months, more than 11 months, more than 1 year or even longer.

In some embodiments, the pharmaceutical compositions of the present disclosure are formulated for parenteral administration, e.g., administered intravenously, subcutaneously or intramuscularly via injection. In some embodiments, the unit dosage for parenteral administration contains one or more compounds provided herein in an amount from about 0.1 mg to about 500 mg of one or more compounds provided herein, for example from about 0.2 mg to about 500 mg, from about 0.3 mg to about 500 mg, from about 0.4 mg to about 500 mg, from about 0.5 mg to about 500 mg, from about 1 mg to about 500 mg, from about 5 mg to about 500 mg, from about 10 mg to about 500 mg, from about 20 mg to about 500 mg, from about 30 mg to about 500 mg, from about 40 mg to about 500 mg, from about 50 mg to about 500 mg, from about 0.5 mg to about 400 mg, from about 0.5 mg to about 300 mg, from about 0.5 mg to about 200 mg, from about 0.5 mg to about 100 mg, from about 0.5 mg to about 90 mg, from about 0.5 mg to about 80 mg, from about 0.5 mg to about 70 mg, from about 0.5 mg to about 60 mg, from about 0.5 mg to about 50 mg, from about 0.5 mg to about 40 mg, from about 1 mg to about 90 mg, from about 5 mg to about 90 mg, from about 10 mg to about 80 mg, from about 20 mg to about 70 mg, from about 30 mg to about 60 mg, or from about 40 mg to about 50 mg, for example about 0.1 mg, about 0.2 mg, about 0.3 mg, about 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg and the like.

In some embodiments, the pharmaceutical composition intended to be administered by injection can be prepared by combining one or more compounds of the present disclosure with sterile, distilled water, sesame or peanut oil, aqueous propylene glycol, so as to form a solution. In some embodiments, the pharmaceutical composition may comprise a surfactant or other solubilizing excipient that is added to facilitate the formation of a homogeneous solution or suspension. In some embodiments, the pharmaceutical composition may further comprise one or more additional agents selected from the group consisting of a wetting agent, a suspending agent, a preservative, a buffer, and an isotonizing agent.

In some embodiments, the pharmaceutical composition intended to be administered by injection can be administered with a syringe. In some embodiments, the syringe is disposable. In some embodiments, the syringe is reusable. In some embodiments, the syringe is pre-filled with the pharmaceutical composition provided herein.

In a further aspect, there is also provided veterinary compositions comprising one or more molecules or compounds of the present disclosure or pharmaceutically acceptable salts thereof and a veterinary carrier. Veterinary carriers are materials useful for the purpose of administering the composition and may be solid, liquid or gaseous materials which are otherwise inert or acceptable in the veterinary art and are compatible with the active ingredient. These veterinary compositions may be administered parenterally, orally or by any other desired route.

The pharmaceutical compositions or veterinary compositions may be packaged in a variety of ways depending upon the method used for administering the drug. For example, an article for distribution can include a container having deposited therein the compositions in an appropriate form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass) , sachets, ampoules, plastic bags, metal cylinders, and the like. The container may also include a tamper-proof assemblage to prevent indiscreet access to the contents of the package. In addition, the container has deposited thereon a label that describes the contents of the container. The label may also include appropriate warnings. The compositions may also be packaged in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water, for injection immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described.

Method of Treatment

In another aspect, the present disclosure provides a method for treating a viral infection in a patient in need thereof, comprising administering an effective amount of any compound described herein.

In some embodiments, the viral infection is from an RNA virus. In certain embodiments, the RNA virus is a single stranded or double stranded RNA virus. In further embodiments, the RNA virus is a positive sense RNA virus or a negative sense RNA virus or an ambisense RNA virus.

In certain embodiments, the viral infection is from a virus selected from the group consisting of Retroviridae virus, Lentiviridae virus, Coronaviridae virus, Picornaviridae virus, Caliciviridae virus, Flaviviridae virus, Togaviridae virus, Bomaviridae, Filoviridae, Paramyxoviridae, Pneumoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Orthomyxoviridae, and Deltavirus.

In certain embodiments, the viral infection is from a virus selected from the group consisting of Lymphocytic choriomeningitis virus, Coronavirus, HIV, SARS, Poliovirus, Rhinovirus, Hepatitis A, Norwalk virus, Yellow fever virus, West Nile virus, Hepatitis C virus, Dengue fever virus, Zika virus, Rubella virus, Ross River virus, Sindbis virus, Chikungunya virus, Boma disease virus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipah virus, Hendra virus, Newcastle disease virus, Human respiratory syncytial virus, Rabies virus, Lassa virus, Hantavirus, Crimean-Congo hemorrhagic fever virus, Influenza and Hepatitis D virus.

In certain embodiments, the viral infection is a coronavirus infection.

In further embodiments, the coronavirus is selected from the group consisting of 229E alpha coronavirus, NL63 alpha coronavirus, OC43 beta coronavirus, HKU1 beta coronavirus, Middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV) , severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) , and SARS-CoV-2.

In certain embodiments, the coronavirus is SARS-CoV-2.

In some embodiments, the viral infection is from a DNA virus. In certain embodiments, the DNA virus is a single stranded or double stranded DNA virus. In further embodiments, the DNA virus is a positive sense DNA virus or a negative sense DNA virus or an ambisense DNA virus.

In certain embodiments, the virus is a Myoviridae, Podoviridae, Siphoviridae, Alloherpesviridae, Herpesviridae (including human herpes virus, and Varicella Zozter virus) , Malocoherpesviridae, Lipothrixviridae, Rudiviridae, Adenoviridae, Ampullaviridae, Ascoviridae, Asfarviridae (including African swine fever virus) , Baculoviridae, Cicaudaviridae, Clavaviridae, Corticoviridae, Fuselloviridae, Globuloviridae, Guttaviridae, Hytrosaviridae, Iridoviridae, Maseilleviridae, Mimiviridae, Nudiviridae, Nimaviridae, Pandoraviridae, Papillomaviridae, Phycodnaviridae, Plasmaviridae, Polydnaviruses, Polyomaviridae (including Simian virus 40, JC virus, BK virus) , Poxviridae (including Cowpox and smallpox) , Sphaerolipoviridae, Tectiviridae, Turriviridae, Dinodnavirus, Salterprovirus, or Rhizidovirus.

In some embodiments, methods described herein may inhibit viral replication transmission, replication, assembly, or release, or minimize expression of viral proteins. In one embodiment, described herein is a method of inhibiting transmission of a virus, a method of inhibiting viral replication, a method of minimizing expression of viral proteins, or a method of inhibiting vims release, comprising administering a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, to a patient suffering from the virus, and/or contacting an effective amount of a compound described herein or a pharmaceutically acceptable salt thereof, with a virally infected cell.

In a further aspect, the present disclosure provides a method for inhibiting RNA polymerase in a subject in need thereof, comprising administrating an effective amount of the compound or a pharmaceutically acceptable salt thereof or the pharmaceutical composition provided herein to the subject.

Combination Therapy

The compounds of the present disclosure can also be used in combination with one or more additional therapeutic or prophylactic agents. As such, also provided herein are methods for treating viral infections in a subject in need thereof, comprising administering to the subject a compound disclosed herein, as a first active ingredient, and a therapeutically effective amount of one or more additional therapeutic or prophylactic agents, as a second active ingredient. Accordingly, there is also provided pharmaceutical compositions comprise one or more compounds of the present disclosure, or a pharmaceutically acceptable salt thereof, as a first active ingredient, and a second active ingredient.

In some embodiments, the second active ingredient include one or more additional therapeutic agents from the same class or group and/or one or ore more additional therapeutic agents from different classes or groups.

In some embodiments, the second active ingredient has complementary activities to the compound provided herein such that they do not adversely affect each other. Such ingredients are suitably present in combination in amounts that are effective for the purpose intended.

In some embodiments, the second active ingredient can be an antibiotic, a protease inhibitor, an anti-viral agent, an anti-inflammatory agent, an immunomodulatory agent, a kinase inhibitor, an anti-metabolite agent, a lysosomotropic agent, a M2 proton channel blocker, a polymerase inhibitor (e.g., EIDD-2801) , a neuraminidase inhibitor, a reverse transcriptase inhibitor, a viral entry inhibitor, an integrase inhibitor, interferons (e.g., types I, II, and III) , or a nucleoside analogue.

In some embodiments, the second active ingredient is an antibiotic. In some embodiments, the antibiotic can be selected from the group consisting of a penicillin antibiotic, a quinolone antibiotic, a tetracycline antibiotic, a macrolide antibiotic, a lincosamide antibiotic, a cephalosporin antibiotic, or an RNA synthetase inhibitor. In some embodiments, the antibiotic is selected from the group consisting of azithromycin, vancomycin, metronidazole, gentamicin, colistin, fidaxomicin, telavancin, oritavancin, dalbavancin, daptomycin, cephalexin, cefuroxime, cefadroxil, cefazolin, cephalothin, cefaclor, cefamandole, cefoxitin, cefprozil, ceftobiprole, cipro, Levaquin, floxin, tequin, avelox, norflox, tetracycline, minocycline, oxytetracycline, doxycycline, amoxicillin, ampicillin, penicillin V, dicloxacillin, carbenicillin, methicillin, ertapenem, doripenem, imipenem/cilastatin, meropenem, amikacin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefoxotin, and streptomycin. In some embodiments, the antibiotic is azithromycin.

In some embodiments, the second active ingredient can be a protease inhibitor. In some embodiments, the protease inhibitor can be selected from the group consisting of nafamostat, camostat, gabexate, epsilon-aminocapronic acid, aprotinin, amprenavir, indinavir, nelfinavir, nirmatrelvir, s-217622, EDP-235, PBI-0451, ritonavir, and saquinavir.

In some embodiments, the second active ingredient can be an anti-viral agent. In some embodiments, the anti-viral agent can be selected from the group consisting of ribavirin, favipiravir, ST-193, oseltamivir, zanamivir, peramivir, danoprevir, ritonavir, remdesivir, cobicistat, elvitegravir, emtricitabine, tenofovir, tenofovir disoproxil, tenofovir alafenamide hemifumarate, abacavir, dolutegravir, efavirenz, elbasvir, ledipasvir, glecaprevir, sofosbuvir, bictegravir, dasabuvir, lamivudine, atazanavir, ombitasvir, lamivudine, stavudine, nevirapine, rilpivirine, paritaprevir, simeprevir, daclatasvir, grazoprevir, pibrentasvir, adefovir, amprenavir, ampligen, aplaviroc, anti-caprine antibody, balavir, cabotegravir, cytarabine, ecoliever, epigallocatechin gallate, etravirine, fostemsavir, gemcitabine, griffithsin, imunovir, indinavir, maraviroc, methisazone, MK-2048, nelfmavir, nevirapine, nitazoxanide, norvir, plerixafor, PRO 140, raltegravir, pyramidine, saquinavir, telbivudine, TNX-355, valacyclovir, VIR-576, ZX-7101A, Baloxavir, Ziresovi, molnupiravir, GS-5806 and zalcitabine.

In some embodiments, the second active ingredient can be an anti-inflammatory agent. In some embodiments, the anti-inflammatory agent can be selected from the group consisting of anti-histamines, corticosteroids, (e.g., fluticasone propionate, fluticasone furoate, beclomethasone dipropionate, budesonide, ciclesonide, mometasone furoate, triamcinolone, flunisolide) , NSAIDs, leukotriene modulators (e.g., montelukast, zafirlukast. pranlukast) , tryptase inhibitors, IKK2 inhibitors, p38 inhibitors, Syk inhibitors, protease inhibitors such as elastase inhibitors, integrin antagonists (e.g., beta-2 integrin antagonists) , adenosine A2a agonists, mediator release inhibitors such as sodium chromoglycate, 5-lipoxygenase inhibitors (zyflo) , DPI antagonists, DP2 antagonists, PI3K delta inhibitors, GGK inhibitors, LP (lysophosphatidic) inhibitors or FLAP (5-lipoxygenase activating protein) inhibitors, bronchodilators (e.g. . muscarinic antagonists, beta-2 agonists) , methotrexate, and similar agents; monoclonal antibody therapy such as anti-lgE, anti-TNF, anti-IL-5, anti-IL-6, anti-IL-12, anti-IL-1 and similar agents; cytokine receptor therapies e.g. etanercept and similar agents; antigen non-specific immunotherapies (e.g. interferon or other cytokines/chemokines, chemokine receptor modulators such as CCR3, CCR4 or CXCR2 antagonists, other cytokine/chemokine agonists or antagonists, TLR agonists and similar agents) , suitable anti-infective agents including antibiotic agents, antifungal agents, anthelmintic agents, antimalarial agents, antiprotozoal agents and antituberculosis agents.

In some embodiments, the second active ingredient can be an immunomodulatory agent. In some embodiments, the immunomodulatory agent can be selected from the group consisting of anti-PD-1 or anti-PDL-1 therapeutics including pembrolizumab, nivolumab, atezolizumab, durvalumab, BMS-936559, or avelumab, anti-TIM3 (anti-HAVcr2) therapeutics including but not limited to TSR-022 or MBG453, anti-LAG3 therapeutics including but not limited to relatlimab, LAG525, or TSR-033, anti-4-1BB (anti-CD37, anti-TNFRSF9) , CD40 agonist therapeutics including but not limited to SGN-40, CP-870, 893 or R07009789, anti-CD47 therapeutics including but not limited to Hu5F9-G4, anti-CD20 therapeutics, anti-CD38 therapeutics, STING agonists including but not limited to ADU-S100, MK-1454, ASA404, or amidobenzimidazoles, anthracyclines including but not limited to doxorubicin or mitoxanthrone, hypomethylating agents including but not limited to azacytidine or decitabine, other immunomodulatory therapeutics including but not limited to epidermal growth factor inhibitors, statins, metformin, angiotensin receptor blockers, thalidomide, lenalidomide, pomalidomide, prednisone, or dexamethasone. In some embodiments, the additional therapeutic agent is a p2-adrenoreceptor agonist including, but not limited to, vilanterol, salmeterol, salbutamol, formoterol, salmefamol, fenoterol carmoterol, etanterol, naminterol, clenbuterol, pirbuterol, flerbuterol, reproterol, bambuterol, indacaterol, terbutaline and salts thereof, for example the xinafoate (1-hydroxy-2-naphthalenecarboxylate) salt of salmeterol, the sulphate salt of salbutamol or the fumarate salt of formoterol.

In some embodiments, the second active ingredient can be a kinase inhibitor. In some embodiments, the kinase inhibitor can be selected from the group consisting of erlotinib, gefitinib, neratinib, afatinib, osimertinib, lapatanib, crizotinib, brigatinib, ceritinib, alectinib, lorlatinib, everolimus, temsirolimus, abemaciclib, LEE011, palbociclib, cabozantinib, sunitinib, pazopanib, sorafenib, regorafenib, sunitinib, axitinib, dasatinib, imatinib, nilotinib, ponatinib, idelalisib, ibrutinib, Loxo 292, larotrectinib, and quizartinib.

EXAMPLES

For the purpose of illustration, the following examples are included. However, it is to be understood that these examples do not limit the present disclosure and are only meant to suggest a method of practicing the present disclosure. Persons skilled in the art will recognize that the chemical reactions described may be readily adapted to prepare a number of other compounds of the present disclosure, and alternative methods for preparing the compounds of the present disclosure are deemed to be within the scope of the present disclosure. For example, the synthesis of non-exemplified compounds according to the present disclosure may be successfully performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by utilizing other suitable reagents and building blocks known in the art other than those described, and/or by making routine modifications of reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present disclosure.

Example 1. Synthesis of Intermediates

1.1 Synthesis of Intermediate 1a:

A solution of compound s-1 (50 g, 0.1718 mol, 1 eq) in TMP (400 mL) was stirred for 15 min at 0 ℃ under N₂ atmosphere. To the above mixture was added POCl₃ (86.18 g, 0.5670 mol, 3.3 eq) dropwise over 30 min at 0 ℃. The resulting mixture was stirred for 1.5 h at 25 ℃ under N₂ atmosphere. The reaction was quenched with ice water at 0 ℃. The resulting mixture was concentrated under vacuum. The concentrated mixture was added acetonitrile (3 L) and filtered to give intermediate 1a (50 g, 0.1348 mol, 78%) as a light-yellow solid. The product was used in the next step directly without further purification.

LCMS: Rt = 0.396 min; m/z= 372 [M+1] ⁺; m/z= 370 [M-1] ^-

¹H NMR (400 MHz, D₂O) δ 8.07 (s, 1H) , 7.35-7.33 (m, 1H) , 7.12-7.11 (m, 1H) , 4.91-4.90 (m, 1H) , 4.50 (d, J = 4.6 Hz, 1H) , 4.41-4.39 (m, 1H) , 4.14-4.10 (m, 2H) .

1.2 Synthesis of Intermediate 1b:

Step 1: Synthesis of compound 1b-2

To a solution of compound 1b-1 (20.0 g, 96.5 mmol, 1.0 eq) in ACN (200 mL) was added DBDMH (15.2 g, 53.1 mmol, 0.55 eq) under N₂ at -30 ℃, the mixture was stirred at -20 ℃ for 2 hrs. Then the reaction mixture was poured into water (200 mL) , extracted with EA (200 mL*3) . The combined organic phase was washed with brine (200 mL) , dried over Na₂SO₄, filtered and evaporated in vacuum to get the crude product. The crude product was purified by FCC (EtOAc in PE from 1%to 15%) to give compound 1b-2 (27.1 g, 98%) as light yellow oil.

LCMS: m/z (M+H) ⁺ =286.0

¹H NMR (400 MHz, CDCl₃) : δ: 7.69 (s, 1H) , 6.90 (d, J = 2.0 Hz, 1H) , 6.76 (d, J = 2.0 Hz, 1H) , 1.50 (s, 9H) .

Step 2: Synthesis of of compound 1b-3

To a solution of compound 1b-2 (20.0 g, 69.9 mmol) in dioxane (50 mL) was added HCl/dioxane (104.8 mL, 4 M, 419.4 mmol) at 0 ℃ under N₂, then the resulting mixture was stirred at 25 ℃ for 5 hrs. The mixture was filtered, and the filter cake was washed with MTBE (100 mL) , then the filtrate was dried in vacuum to give compound 1b-3 (11.3 g, 50.8 mmol, 72.7%) as a white solid. The crude product was used directly for next step without further purification.

LCMS: m/z =186.0

¹H NMR (400 MHz, DMSO-d₆) : δ ppm = 7.24 (d, J = 2.0 Hz, 1H) , 6.92 (d, J = 2.0 Hz, 1H) .

Step 3: Synthesis of compound 1b-4

To a solution of compound 1b-3 (16.0 g, 71.9 mmol) in EtOH (200 mL) was added formamidine acetate (37.4 g, 359.6 mmol) and K₃PO₄ (76.3 g, 359.6 mmol) at 25 ℃ under N₂, then the mixture was stirred at 78 ℃ for 16 hrs. The reaction mixture was filtered and the filter cake was washed with EtOH (100 mL*2) , then the filtrate was evaporated in vacuum to get the crude product. The crude product was triturated by THF: DCM (1: 3) and filtered. The filter cake was dried in vacuum, and the filtrate was further purified by FCC (10～26%THF in PE) to give compound 1b-4 (9.5 g, 44.6 mmol, 62.0%) as light yellow solid.

LCMS: m/z = 213.0

¹H NMR (400 MHz, DMSO-d₆) : δ ppm = 7.84 (br, 2H) , 7.82 (s, 1H) , 7.81 (d, J = 1.6 Hz, 1H) , 6.97 (d, J = 1.6 Hz, 1H) .

Step 4: Synthesis of compound 1b-5

To a solution of compound 1b-4 (21.1 g, 99.0 mmol) in THF (200 mL) was added D₂O (10 mL) , then it was concentrated to dryness three times. After that, THF (200 mL) and D₂O (10 mL) was added into the residue under N₂, then the solution was added Pd (dppf) Cl₂ (7.2 g, 9.9 mmol) and TMEDA (2.3 g, 19.8 mmol) at 25 ℃. The reaction was stirred at 25 ℃ for 0.5 hour. After that, the solution was cooled to 0 ℃, NaBD₄ (8.3 g, 198.0 mmol) was added into the solution, the reaction was stirred at 25 ℃ for 17 hours. TLC (PE: EA = 1: 1) showed compound 1b-4 was consumed completely and a new spot was detected. The mixture was poured into saturated NH₄Cl (200 mL) . The reaction mixture was extracted with EA (100 mL x 3) . The organic phase was washed with brine (100 mL) , then dried over Na₂SO₄ and concentrated to give the residue. The residue was purified by column chromatography on silica gel (EtOAc in PE from 10%to 50%) to give compound 1b-5 (7.5 g, 56.3%) as yellow solid.

LCMS: m/z = 136.2

¹H NMR (400 MHz, DMSO-d₆) : δ: 7.80 (s, 1H) , 7.70 (s, 2H) , 7.61 (d, J = 1.6 Hz, 1H) , 6.86 (d, J = 1.6 Hz, 1H) .

Step 5: Synthesis of Compound 1b-6

A solution of compound 1b-5 (4.0 g, 29.5 mmol, 1.0 eq) in DMF (36 mL) was cooled to -5℃～5℃. NIS (6.7 g, 29.5 mmol, 1.0 eq) was added to the above reaction under N₂ atmosphere. The reaction was stirred at -5℃～5℃ for 2 h. TLC showed the reaction was finished. The reaction mixture was poured in to aqueous NaOH (400 mL, 1M) and stirred for another 30 min. The solid crushed out was filtered off and the solid was washed with H₂O (50 mL) , PE (60 mL) , dried in vacuo to give compound 1b-6 as a solid (6 g, 77%) .

¹H NMR (400 MHz, DMSO-d₆) δ: 7.87 (s, 1H) , 7.80 (br s, 2H) , 6.86 (s, 1H) .

Step 6: Synthesis of Compound 1b-7

To a solution of compound 1b-6 (2.6 g, 9.9 mmol, 1.0 eq) in THF (40 mL) was added TMSCl (2.4 g, 21.7 mmol, 2.2 eq) under N₂ atmosphere and the reaction was stirred at rt for 40 min. The reaction mixture was cooled to -5～5℃, PhMgCl (11.8 mL, 2M, 23.6 mmol) was added to the above reaction mixture and keep the temperature at -5～5℃ for another 1.5 h. Then i-PrMgCl. LiCl (9.9 mL, 12.9 mmol, 1.3eq) was added to the above reaction mixture and stirred for another 15 min after addition. A solution of compound A (4.1 g, 9.9 mmol, 1.0 eq) in THF (40 mL) was added to the above reaction mixture at -20～-15℃. After addition, the reaction mixture was warmed to rt and stirred for another 1 h. TLC (PE: EA=1: 1) showed the reaction was finished, 1M HCl was added to the reaction to modify pH=2～3. EtOAc was added, the two phases were separated, and the organic phase was washed with HCl (1M) twice. Then the organic phase was washed with saturated NaHCO₃, brine, dried over Na₂SO₄, filtered and concentrated in vacuo to get a crude product, which was purified by column (PE: EA=2: 1) to give compound 1b-7 (2.0 g) as solid.

Step 7: Synthesis of Compound 1b-8

To a solution of compound 1b-7 (2.0 g, 3.6 mmol, 1.0 eq) in DCM (24 mL) was added TfOH (1.1 g, 7.2 mmol) at -78℃. After 10 min, TMSOTf (1.7 g, 7.6 mmol, 2.1eq) was added to the above reaction and stirred for another 30 min. TMSCN (1.4 g, 14.4 mmol, 4.0 eq) was then added. After 10 min, TLC (PE: EA=1: 1) showed the reaction was finished. TEA was then added to quench the reaction, followed by the addition of NaHCO₃ to pH ～8. The two phases were separate and the aqueous phase was extracted with DCM. The combined organic phase was dried over Na₂SO₄, filtered and concentrated in vacuo to get a crude product, which was purified by column to give compound 1b-8 (1.4 g, 69%) as solid.

Step 8: Synthesis of Compound 1b-9

To a solution of compound 1b-8 (1.6 g, 2.8 mmol, 1.0 eq) in DCM (23 mL) was added BCl₃ (11.1 mL, 11.1 M) at-78℃. The reaction mixture was stirred at -40 ℃ for 2 h. MeOH (10 mL) was added to quench the reaction, followed by the addition of TEA to pH 8～9. The solution was concentrated. Then water (100 mL) was added, the aqueous phase was washed with DCM, and the aqueous phase was concentrated to about 50 mL. Another 100 mL was added and repeat the same procedure seven times. The solid formed was filtered and concentrated in vacuo to give compound 1b-9 (0.4 g, 51%) as solid.

¹H NMR (400 MHz, DMSO-d₆) δ: 7.9-787 (m, 3H) , 6.86 (s, 1H) , 6.07 (s, 1H) , 5.15 (s, 1H) , 4.90-4.87 (m, 1H) , 4.62-4.59 (m, 1H) , 4.03-4.01 (m , 1 H) , 3.94-3.90 (m, 1H) , 3.63-3.46 (m, 2H) .

Step 9: Synthesis of Compound 1b:

A solution of compound 1b-9 (50.0 g, 0.1718 mol, 1 eq) in TMP (400 mL) was stirred for 15 min at 0 ℃ under N₂ atmosphere. To the above mixture was added POCl₃ (86.2 g, 56.7 mmol, 3.3 eq) dropwise over 30 min at 0 ℃. The resulting mixture was stirred for 1.5 h at 25 ℃ under N₂ atmosphere. The reaction was quenched with ice water at 0 ℃. The resulting mixture was concentrated under vacuum. The crude product was precipitated by the addition of acetonitrile (3 L) . The precipitated solids were collected by filtration to give compound 1b (50.0 g, 134.8 mmol, 78%) as a light-yellow solid.

LCMS: m/z= 373

Example 2. Synthesis of Compounds

2.1 Synthesis of compound 51

Step 1: Synthesis of compound 51-2:

To a mixture of compound 51-1 (8 g, 23.36 mmol, 1.0 eq) in cyclohexane (65 mL) was added Py (3.7 g, 46.72 mmol, 2.0 eq) , followed by addition of compound s-2 (3.61 g, 28.03 mmol, 1.2 eq) dropwise, then the mixture was stirred at r.t for 4 h. The reaction mixture was quenched with NH₄Cl (aq) and extracted with PE (50 mL x 3) , the combined organic layer was washed with brine (50 mL x 3) , dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by FCC (n-hexane=100%) to obtained compound 51-2 (6.6 g, white solid) .

Step 2: Synthesis of compound 51:

To a mixture of Intermediate 1a (1g, 0.0027 mol, 1.0 eq) in DMF (15 mL) were added compound 51-2 (1.36 g, 0.004 mol, 1.5 eq) and DIEA (2.09 g, 0.0162 mol, 6 eq) . The reaction mixture was stirred at 80 ℃ for 16 h. The reaction mixture was directly purified by Prep-HPLC to give compound 51 (122 mg) .

LCMS: Rt = 1.956 min; m/z= 670 [M+1] ⁺; m/z= 668 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.93 (s, 1H) , 7.06-7.05 (m, 2H) , 5.45 -5.42 (m, 2H) , 4.38-4, 25 (m, 1H) , 4.25-4.20 (m, 1H) , 4.20-4.15 (m, 1H) , 4.15-4.06 (m, 2H) , 4.05-4.00 (m, 1H) , 1.70-1.50 (m, 2H) , 1.30-1.20 (m, 26H) , 0.88 (t, J = 6.8 Hz, 3H) .

The following compounds were synthesized according to the same procedure as for compound 51 except for using different starting materials or intermediates:

Compound 7:

LCMS: m/z= 671.30 [M+1] ⁺; m/z= 669.30 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.94 (s, 1H) , 7.06 (s, 1H) , 5.45 -5.40 (m, 2H) , 4.83-4.80 (m, 1H) , 4.33-4.28 (m, 1H) , 4.23 (t, J = 5.3 Hz, 1H) , 4.16-4.11 (m, 1H) , 4.07-4.06 (m, 2H) , 4.05-3.99 (m, 1H) , 1.61-1.54 (m, 2H) , 1.27-1.25 (m, 26H) , 0.897 (t, J = 6.7 Hz, 3H) .

Compound 8:

LCMS: m/z= 699.40 [M+1] ⁺; m/z= 697.35 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.94 (s, 1H) , 7.06 (s, 1H) , 5.45-5.40 (m, 2H) , 4.81-4.79 (m, 1H) , 4.33-4.28 (m, 1H) , 4.23 (t, J = 5.3 Hz, 1H) , 4.16-4.11 (m, 1H) , 4.07-3.99 (m, 3H) , 1.61-1.54 (m, 2H) , 1.27-1.25 (m, 30H) , 0.87 (t, J = 6.7 Hz, 3H) .

Compound 19:

LCMS: m/z=572.40 [M+1] ⁺; m/z= 570.10 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.95 (s, 1H) , 7.31-7.08 (m, 2H) , 5.45-5.42 (m, 2H) , 4.85-4.77 (m, 1H) , 4.71-4.67 (m, 1H) , 4.25-4.23 (m, 2H) , 4.13-4.04 (m, 2H) , 1.56-1.30 (m, 2H) , 1.35-1.21 (m, 10H) , 1.21-1.20 (m, 3H) , 0.89 (t, J = 6.6 Hz, 3H) .

Compound 23:

LCMS: m/z= 684.55 [M+1] ⁺; m/z= 682.25 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 7.95 (s, 1H) , 7.11-7.08 (m, 2H) , 5.45-5.42 (m, 2H) , 4.87-4.83 (m, 1H) , 4.71-4.67 (m, 1H) , 4.25-4.23 (m, 2H) , 4.13-4.04 (m, 2H) , 1.56-1.30 (m, 2H) , 1.35-1.21 (m, 29H) , 0.89 (t, J = 6.6 Hz, 3H) .

Compound 53:

LCMS: m/z= 698.55 [M+1] ⁺; m/z= 696.30 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.92 (s, 1H) , 7.05 (d, J = 4.7 Hz, 1H) , 7.01 (d, J = 4.7 Hz, 1H) , 5.46-5.38 (m, 2H) , 4.81-4.79 (m, 1H) , 4.33-4.30 (m, 1H) , 4.23 (t, J = 5.2 Hz, 1H) , 4.17-4.11 (m, 1H) , 4.09-4.06 (m, 2H) , 4.05-3.99 (m, 1H) , 1.62-1.54 (m, 2H) , 1.36-1.33 (m, 2H) , 1.27-1.23 (m, 28H) , 0.88 (t, J = 6.8 Hz, 3H) .

Compound 62:

LCMS: [MS+1] ⁺ = 656.45, [MS-1] ^-= 654.15.

¹H NMR (400 MHz, CD₃OD) δ 8.02 (s, 1H) , 7.20 (d, J = 4.3 Hz, 1H) , 7.14 (d, J = 4.3 Hz, 1H) , 5.52-5.48 (m, 2H) , 4.80 (d, J = 5.1 Hz, 1H) , 4.73-4.64 (m, 1H) , 4.39-4.30 (m, 1H) , 4.25 (t, J = 5.3 Hz, 1H) , 4.23-4.15 (m, 1H) , 4.14-4.03 (m, 1H) , 1.66-1.46 (m, 4H) , 1.28-1.24 (m, 20H) , 0.96-0.79 (m, 6H) .

Compound 127:

LCMS: m/z= 685.30 [M+1] ⁺; m/z= 683.30 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 7.86 (s, 1H) , 6.88 (s, 1H) , 5.44-5.40 (m, 2H) , 4.82 (d, J = 5.1 Hz, 1H) , 4.69-4.68 (m 1H) , 4.32 (d, J = 4.3 Hz, 1H) , 4.23 (t, J = 5.1 Hz, 1H) , 4.15-4.09 (m, 1H) , 4.05-4.00 (m, 1H) , 1.57-1.51 (m, 2H) , 1.28-1.20 (m, 26H) , 1.19 (d, J = 6.7 Hz, 3H) , 0.88 (t, J = 6.7 Hz, 3H) .

Compound 128:

LCMS: [MS+1] ⁺ = 685.35, [MS-1] ^-= 683.30

¹H NMR (400 MHz, CD₃OD) δ 7.91 (s, 1H) , 6.97 (s, 1H) , 5.45 (dd, J = 11.5, 2.3 Hz, 2H) , 4.84 (d, J = 5.2 Hz, 1H) , 4.74-4.69 (m, 1H) , 4.36 -4.33 (m, 1H) , 4.26 (t, J = 5.2 Hz, 1H) , 4.19-4.13 (m, 1H) , 4.07-4.01 (m, 1H) , 1.59-1.44 (m, 2H) , 1.29-1.25 (m, 26H) , 1.22 (d, J = 6.3 Hz, 3H) , 0.90 (t, J = 6.8 Hz, 3H) .

2.2 Synthesis of compound 57:

Step 1: Synthesis of compound 57-2:

To a mixture of compound 57-1 (1 g, 3.06 mmol, 1eq) in cyclohexane (20 mL) was added compound s-2 (473 mg, 3.67 mmol, 1.2 eq) and py (484 mg, 6.12 mmol, 2 eq) , then the mixture was stirred at r. t for 4 h. The reaction mixture was quenched with NH₄Cl (aq) and extracted with PE (50 mL x 3) , the combined organic layer was washed with brine (50 mL x 3) , dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by FCC to obtained compound 57- 2 (0.98 g) .

¹H NMR (400 MHz, CDCl₃) δ 5.71 (s, 2H) , 4.22-4.18 (m, 2H) , 1.69-1.64 (m, 2H) , 1.24 (br s, 38 H) , 0.86 (t, J = 6.6 Hz, 3H) .

Step 2: Synthesis of compound 57-3:

To a solution of compound 57-2 (0.98 g, 2.34 mmol, 1.0 eq) in acetone (10 mL) was added NaHCO₃ (0.59 g, 7.03 mmol, 3 eq. ) and NaI (1.4 g, 9.38 mmol, 4 eq) , the mixture was stirred under N₂ at 45 ℃ for 4 h. The reaction mixture was quenched with H₂O (aq) (30 mL) , and extracted with PE (30 mL x 3) , the combined organic layer was washed with brine (40 mL x 3) , dried over Na₂SO₄, filtered and concentrated to give the compound 57-3 (1.16 g) .

Step 3: Synthesis of compound 57:

To a mixture of Intermediate 1a (300 mg, 0.800 mmol, 1.0 eq) in DMF (20 mL) were added DIEA (521 mg, 4.043 mmol 5 eq) and compound 57-3 (1.237 g, 2.426 mmol, 3 eq) . The reaction mixture was stirred at 45 ℃ for 16 h. The reaction mixture was directly purified with column (DCM: MeOH=5: 1) . The organic layer was concentrated under reduced pressure. The resulting residue was purified again with prep-TLC (DCM: MeOH=5: 1) to give compound 57 (23.66 mg) .

LCMS: Rt = 2.2 min; m/z= 754 [M+1] ⁺; m/z= 752 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 7.85 (s, 1H) , 6.97 (d, J = 4.6 Hz, 1H) , 6.86 (d, J = 4.6 Hz, 1H) , 5.43 (d, J = 13.0 Hz, 2H) , 4.85-4.81 (m, 1H) , 4.32-4, 31 (m, 1H) , 4.24-4.23 (m, 1H) , 4.12-4.00 (m, 3H) , 1.59-1.57 (m, 2H) , 1.38-1.35 (m, 8H) , 1.26 (br s, 31H) , 0.88 (t, J = 6.8 Hz, 3H) .

2.3 Synthesis of compound 85:

Step 1: Synthesis of compound 85-2:

To a mixture of compound 85-1 (1 g, 2.94 mmol, 1eq) in DCM: H₂O (20 mL 1: 1) was added K₂CO₃ (1.54 g, 11.2 mmol, 3.8 eq) , (nBu₄N) ₂SO₄ (170 mg, 0.29 mmol, 0.1 eq) and compound s-3 (532 mg, 3.23 mmol, 1.1 eq) , then the mixture was stirred at r. t for 16 h. The reaction mixture was quenched with H₂O and extracted with PE (50 mL x 3) , the combined organic layer was washed with brine (50 mL x 2) , dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by FCC to obtain compound 85-2 (0.9 g) .

Step 2: Synthesis of compound 85:

To a mixture of Intermediate 1a (300 mg, 0.800 mmol, 1.0 eq) in DMF (20 mL) were then added DIEA (521 mg, 4.043 mmol, 5 eq) and compound 85-2 (1.02 g, 2.426 mmol, 3 eq) . The reaction mixture was stirred at 45 ℃ for 16 h. The reaction mixture was purified with column (DCM: MeOH=5: 1) . The organic layer was concentrated under reduced pressure and further purified by prep-TLC (DCM: MeOH=5: 1) to give compound 85 (21.9 mg) .

m/z= 724 [M+1] ⁺; m/z= 722 [M-1] ^-

¹H NMR (400 MHz, DMSO-d₆) δ 8.00-7.75 (m, 2H) , 6.85 (d, J = 4.5 Hz, 1H) , 6.80 (d, J = 4.3 Hz, 1H) , 6.12 (d, J = 5.9 Hz, 1H) , 5.50 (s, 1H) , 5.28 (d, J = 12.5 Hz, 2H) , 4.55 (s, 1H) , 4.06 (s, 1H) , 3.85 (m, 2H) , 3.70 (s, 1H) , 3.56 -3.48 (m, 1H) , 3.06 -3.00 (m, 1H) , 2.22-2.18 (m , 2H) , 1.42 (br s, 2H) , 1.19 (br s, 36H) , 0.81 (t, J=6.8 Hz, 3H) .

The following compounds were synthesized according to the same procedure as for compound 85 except for using different starting materials or intermediates:

Compound 75:

LCMS: m/z = 584.40 [M+1] ⁺; m/z = 582.20 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 8.01 (s, 1H) , 7.24 (d, J = 4.5 Hz, 1H) , 7.15 (d, J = 4.7 Hz, 1H) , 5.48-5.39 (m, 2H) , 4.77-4.75 (m, 1H) , 4.34-4.29 (m, 1H) , 4.22 (t, J = 5.5 Hz, 1H) , 4.18-4.12 (m, 1H) , 4.08-4.00 (m, 1H) , 2.30-2.25 (m, 2H) , 1.54 (t, J = 7.5 Hz, 2H) , 1.27-1.23 (m, 16H) , 0.87 (t, J = 6.6 Hz, 3H) .

Compound 79:

LCMS: m/z = 640.45 [M+1] ⁺; m/z = 638.20 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.97 (s, 1H) , 7.14 (d, J = 4.7 Hz, 1H) , 7.08 (d, J = 5.9 Hz, 1H) , 5.46-5.42 (m, 2H) , 4.80-4.76 (m, 1H) , 4.34-4.29 (m, 1H) , 4.23-4.18 (m, 1H) , 4.17-4.12 (m 1H) , 4.06-4.01 (m, 1H) , 2.31-2.22 (m, 2H) , 1.57-1.53 (m, 2H) , 1.26 (br s, 24H) , 0.89 (t, J=6.8 Hz, 3H) .

Compound 81:

LCMS: [MS+1] ⁺ = 668.4, [MS-1] ^-= 666.15

¹H NMR (400 MHz, CD₃OD) δ 8.00 (s, 1H) , 7.15 (d, J = 4.8 Hz, 1H) , 7.08 (d, J = 4.8 Hz, 1H) , 5.49-5.47 (m, 2H) , 4.79-4.77 (d, J = 5.5 Hz, 1H) , 4.38 -4.29 (m, 1H) , 4.24 -4.14 (m, 2H) , 4.12 -4.03 (m, 1H) , 2.34 -2.27 (m, 2H) , 1.63 -1.55 (m, 2H) , 1.31 -1.27 (m, 28H) , 0.91 (t, J = 6.8 Hz, 3H) .

Compound 83:

LCMS: [MS+1] ⁺ = 696.50, [MS-1] ^-= 694.25.

¹H NMR (400 MHz, CD₃OD) δ 7.87 (s, 1H) , 6.97 (d, J = 4.6 Hz, 1H) , 6.86 (d, J = 4.6 Hz, 1H) , 5.44 (d, J=12 Hz, 2H) , 4.80 (d, J = 5.2 Hz, 1H) , 4.32-4.22 (m, 2H) , 4.16-3.99 (m, 2H) , 2.27 (t, J = 7.5 Hz, 2H) , 1.59-1.47 (m, 2H) , 1.26-1.22 (m, 32H) , 0.87 (t, J = 6.7 Hz, 3H) .

2.4 Synthesis of compound 66:

Step 1: Synthesis of compound 66-2:

To a mixture of compound 66-1 (8 g, 17.75 mmol, 1eq) in THF (480 mL) was added LiAlH₄ (2.5 M, 10.65 mL, 1.5 eq) dropwise at rt, then the mixture was stirred at rt for 1 h. The reaction mixture was quenched with potassium sodium tartrate (solution) and extracted with PE (150 mL x 3) . The combined organic layer was washed with brine (50 mL x 3) , dried over Na₂SO₄, filtered and concentrated to afford compound 66-2 as white solid, which was used directly for next step.

¹H NMR (400 MHz, CDCl₃) δ 3.56-3.54 (m, 1H) , 1.44-1.37 (m, 4H) , 1.34-1.14 (m, 52H) , 0.86 (t, J = 6.6 Hz, 6H) .

Step 2: Synthesis of compound 66-4:

To a mixture of compound 66-2 (12.64 g, 27.91 mmol, 1eq) in cyclohexane: DCM (120 mL: 12 mL) was added compound 66-3 (14.4 g, 111.65 mmol, 4.0 eq) and pyridine (13.25 g, 167.49 mmol, 6.0 eq) at 0℃, then the mixture was stirred at rt for 4 h. The reaction mixture was quenched with saturated NH₄Cl and extracted with ethyl acetate (70 mL x 3) . The combined organic layer was washed with brine (50 mL x 3) , dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by FCC (PE=100%) to obtained compound 66-4 (7.98 g, 52%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 5.71 (s, 2H) , 4.78-4.76 (m, 1H) , 1.66-1.54 (m, 4H) , 1.28-1.22 (m, 52H) , 0.86 (t, J = 6.8 Hz, 6H) .

Synthesis of compound 66-5:

To a mixture of compound 66-4 (2.98 g, 5.46 mmol, 1.0 eq) in acetone (30 mL) was added NaI (4.1 g, 27.32 mmol, 5.0 eq) and NaHCO₃ (1.84 g, 21.86 mmol, 4.0 eq) . Then the mixture was stirred at 45℃ for 16 h. The reaction mixture was filtered and the filtrate was concentrated to dryness. The residue was purified by FCC (PE =100%) to obtained compound 66-5 (2.2 g, 63%) as a yellow solid.

¹H NMR (400 MHz, CDCl₃) δ 5.93 (s, 2H) , 4.77-4.74 (m, 1H) , 1.58-1.53 (m, 4H) , 1.28-1.22 (m, 52H) , 0.86 (t, J = 6.6 Hz, 6H) .

Synthesis of compound 66:

To a mixture of Intermediate 1a (332 mg, 0.90 mmol, 1.0 eq) in DMF (25 mL) was added DIEA (577 mg, 4.48 mmol, 5 eq) and compound 66-5 (1.71 g, 2.7 mmol, 3 eq) . The reaction mixture was stirred at 45℃ for 24 h. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash column on silica gel (DCM: MeOH=80: 20) to afford a crude product, which was purified by prep-TLC to afford compound 66 (29 mg, 4%) .

¹H NMR (400 MHz, CD₃OD) δ 7.85 (s, 1H) , 6.96 (d, J = 4.6 Hz, 1H) , 6.87 (d, J = 4.6 Hz, 1H) , 5.50-5.41 (m, 2H) , 4.68-4.62 (m, 2H) , 4.34-4.32 (m, 1H) , 4.25 (t, J = 5.1 Hz, 1H) , 4.16-4.10 (m, 1H) , 4.04-3.99 (m, 1H) , 1.59-1.48 (m, 4H) , 1.37-1.12 (d, J = 10.7 Hz, 52H) , 0.87 (t, J = 6.7 Hz, 6H) .

LCMS: m/z= 880.55 [M+1] ⁺

Compound 24 was synthesized according to the similar procedure as mentioned above.

Compound 24:

LCMS: [MS+1] ⁺ =712.55, [MS-1] ^-=710.25

¹H NMR (400 MHz, CD3OD) δ 7.85 (s, 1H) , 6.96 (d, J = 4.6 Hz, 1H) , 6.88 (d, J = 4.5 Hz, 1H) , 5.47-5.43 (m, 2H) , 4.85-4.80 (m, 1H) , 4.75-4.60 (m, 1H) , 4.33-4.31 (m, 1H) , 4.17-4.15 (m, 2H) , 4.04-4.02 (m, 1H) , 1.56-1.26 (m, 2H) , 1.24-1.19 (m, 33H) , 0.88 (t, J = 6.6 Hz, 3H) .

2.5 Synthesis of compound 125:

Step 1: Synthesis of compound 125-3:

To a solution of Mg (3.12 g, 1.4 eq) in THF (250 mL) was added I₂ (228.8 mg, 0.01 eq) and 1-bromotetradecane (25 g, 90.16 mmol, 26.88 mL, 1 eq) was added slowly at 50 ℃ under N₂. The mixture was stirred at 50 ℃ for 5 hrs. The solution was turned to cloudy gray color. To a solution with a cloudy gray color of compound 125-2 (above-mentioned) was added Li₂CuCl₄ (0.1 M, 18.60 mL) and (2R) -2-methyloxirane (3 g, 1 eq) slowly at -78℃. The reaction mixture was stirred at -78℃ for 0.5 hr, then an additional 2 hrs at 25℃. The mixture was quenched by aq NH₄Cl (1L) at 0℃ and extracted with EA (1L*3) . The organic layer was washed with brine, dried over Na₂SO₄ and concentrated in vacuo. The residue was purified by flash silica gel chromatography (330 gSilica Flash Column, Eluent of 0～10%Ethyl acetate/Petroleum ether gradient @100 mL/min) to obtain compound 125-3 (14 g, 52.8%yield) as a white solid.

¹H NMR (400MHz, CDCl₃) δ 3.85-3.80 (m, 1H) , 1.45-1.41 (m, 2H) , 1.29-1.25 (m, 26H) , 1.21 (d, J = 6.2 Hz, 3H) , 0.90 (t, J = 6.8 Hz, 3H) .

Synthesis of compound 125-5:

To a mixture of compound 125-3 (2 g, 0.0078 mol, 1.0 eq) in cyclohexane (50 mL) was added pyridine (1.24 g, 0.0156 mol, 2.0 eq) , followed by compound 125-4 (1.21 g, 0.0094 mol, 1.2 eq) dropwise_, then the mixture was stirred at r. t for 16 h. The reaction mixture was quenched with ice-water and extracted with ethyl acetate (150 mL x 3) , the combined organic layer was washed with brine (70 mL x 3) , dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by FCC (n-hexane=100%) to obtained compound 125-5 (1.6 g, 60%yield) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 5.70 (s, 2H) , 4.82-4.80 (m, 1H) , 1.67-1.65 (m, 2H) , 1.35-1.23 (m, 29H) , 0.87 (t, J = 6.6 Hz, 3H) .

Synthesis of compound 125:

To a mixture of intermediate 1a (500 mg, 1.38 mmol, 1.0 eq) in DMF (8 mL) was added compound 125-5 (940 mg, 2.7 mmol, 2 eq) and DIEA (1.07 g, 8.28 mmol, 6 eq) . The reaction mixture was stirred at 80 ℃ for 16 h. The reaction mixture was directly purified by prep-HPLC to give compound 125 (44.3 mg, 4.7%) .

LCMS: m/z= 684.35 [M+1] ⁺; m/z= 682.25 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 7.95 (s, 1H) , 7.11-7.08 (m, 2H) , 5.45-5.42 (m, 2H) , 4.80-4.79 (m, 1H) , 4.71-4.67 (m, 1H) , 4.25-4.23 (m, 2H) , 4.13-4.04 (m, 2H) , 1.56-1.30 (m, 2H) , 1.35-1.21 (m, 26H) , 1.21-1.20 (m, 3H) , 0.89 (t, J = 6.6 Hz, 3H) .

The following compounds were synthesized according to the same procedure as for compound 125 except for using different starting materials or intemediates:

Compound 126:

LCMS: m/z= 684.30 [M+1] ⁺; m/z= 682.30 [M-1] ^-

¹H NMR (400 MHz, CD3OD) δ 7.93 (s, 1H) , 7.07-7.04 (m, 2H) , 5.46-5.40 (m, 2H) , 4.80 (d, J = 5.1 Hz, 1H) , 4.69 (q, J = 6.3 Hz, 1H) , 4.32 (t, J = 4.2 Hz, 1H) , 4.24 (t, J = 5.2 Hz, 1H) , 4.17-4.11 (m, 1H) , 4.06-4.00 (m, 1H) , 1.62-1.40 (m, 2H) , 1.32-1.23 (m, 26H) , 1.20 (d, J = 6.2 Hz, 3H) , 0.88 (t, J = 6.8 Hz, 3H) .

Compound 129:

LCMS: m/z=712.35 [M+1] ⁺; m/z= 710.25 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 7.93 (s, 1H) , 7.05-7.04 (m, 2H) , 5.48-5.40 (m, 2H) , 4.80 (d, J = 5.2 Hz, 1H) , 4.71 -4.66 (m, 1H) , 4.35-4.30 (m, 1H) , 4.25 (t, J = 5.2 Hz, 1H) , 4.14-4.12 (m, 1H) , 4.04-4.03 (m, 1H) , 1.65-1.41 (m, 2H) , 1.29-1.23 (m, 30H) , 1.21 (d, J = 6.2 Hz, 3H) , 0.92 -0.85 (t, J = 6.2 Hz, 3H) .

Compound 130:

LCMS: m/z = 712.45 [M+1] ⁺; m/z = 710.40 [M-1] ^-

¹H NMR (400 MHz, CD₃OD) δ 7.98 (s, 1H) , 7.17 (d, J = 4.7 Hz, 1H) , 7.12 (d, J = 4.7 Hz, 1H) , 5.48-5.40 (m, 2H) , 4.80 (d, J = 5.2 Hz, 1H) , 4.71-4.66 (m, 1H) , 4.35-4.30 (m, 1H) , 4.25-4.12 (m, 2H) , 4.06-4.01 (m, 1H) , 1.65-1.41 (m, 2H) , 1.29-1.23 (m, 30H) , 1.21 (d, J = 6.2 Hz, 3H) , 0.89 (t, J = 6.2 Hz, 3H) .

2.6 Synthesis of compound 117:

Step 1: Synthesis of compound 117-3

A solution of NaH (4.5 g, 113.5 mmol, 60%in mineral oil) , Diisopropylamine (16.0 mL, 113.5 mmol) in THF (100 mL) was degassed with N₂ for three times, then compound 117-1 (10.5 mL, 113.5 mmol) was added dropwise. After stirring at 70 ℃ for 30 min, the mixture was cooled to 0 ℃. Then n-BuLi (45.4 mL, 2.5M) was added dropwise, the mixture was stirred at 0 ℃ for 2 hours. Then compound 117-2 (35.8 mL, 113.5 mmol) was added dropwise, the resulting mixture was stirred at 25 ℃ for 16 hours. The mixture was quenched with H₂O (20 mL) , extracted with EtOAc (100 mL × 3) . The combined organic phase was dried over Na₂SO₄, filtered and concentrated to get a residue, which was purified by column to afford compound 117-3 (35.0 g, 98%yield) as white solid.

¹H NMR (400 MHz, CD₃OD) δ: 1.47-1.41 (m, 2H) , 1.34-1.24 (m, 28H) , 1.09 (s, 6H) , 0.90 (t, J=6.8 Hz, 3H) .

Step 2: Synthesis of compound 117-5

A solution of compound 117-3 (0.5 g, 5.7 mmol) , K₂CO₃ (3.2 g, 23.0 mmol) , compound 117-4 (195 mg, 0.6 mmol) in DCM (6 mL) and H₂O (1.5 mL) was degassed with N₂ for three times. After stirring at 25 ℃ for 20 min, a solution of compound 117-4 (947 mg, 5.7 mmol) in DCM (0.6 mL) was added dropwise, the resulting mixture was stirred at 25 ℃ for 16 hours. The mixture was extracted with DCM (50 mL × 3) . The combined organic layer was washed with brine (30 mL) , dried over anhydrous Na₂SO₄, filtrated and concentrated to afford the residue, which was purified by flash column chromatography (0%to 3%EtOAc in PE) to afford compound 117-5 (230 mg, 30%yield) as a colorless oil.

¹H NMR (400 MHz, CDCl₃) δ ppm 5.71 (s, 2H) , 1.55-1.46 (m, 2H) , 1.32-1.22 (m, 28H) , 1.19 (s, 6H) , 0.88 (t, J = 6.4 Hz, 3H) .

Step 3: Synthesis of compound 117

A solution of intermediate 1a (360 mg, 1.0 mmol) , DIEA (1.0 mL, 6.0 mmol) in DMF (5 mL) was added compound 117-5 (350 mg, 1.0 mmol) , the mixture was stirred at 80 ℃ for 48 hours. The mixture was concentrated to afford the residue, which was purified by prep-HPLC (0.1%NH₄HCO₃) to afford compound 117 (77.7 mg, 12%yield) .

LCMS: m/z (M+H) ⁺ = 696.6; (M-H) ^-= 694.5

¹H NMR (400 MHz, DMSO-d₆) δ: 8.03-7.71 (m, 3H) , 6.92 (d, J = 4.6 Hz, 1H) , 6.80 (d, J = 4.6 Hz, 1H) , 6.23-6.11 (m, 1H) , 5.37 (d, J = 12.4Hz, 2H) , 4.65-4.56 (m, 1H) , 4.18-4.09 (m, 1H) , 3.99-3.89 (m, 2H) , 3.89-3.81 (m, 1H) , 1.47-1.35 (m, 2H) , 1.29-1.11 (m, 28H) , 1.07 (s, 6H) , 0.85 (t, J=6.8 Hz, 3H) .

Compound 131 was synthesized according to the same procedure as for compound 117 except for using different starting materials or intermediates.

Compound 131:

LCMS: [MS+1] ⁺ = 724.35, [MS-1] ^-= 722.30

¹H NMR (400 MHz, CD₃OD) δ 7.93 (s, 1H) , 7.06 (d, J = 4.7 Hz, 1H) , 7.03 (d, J = 4.7 Hz, 1H) , 5.45 (d, J = 11.6 Hz, 2H) , 4.82 (d, J = 5.2 Hz, 1H) , 4.38-4.32 (m, 1H) , 4.25 (t, J = 5.3 Hz, 1H) , 4.20-4.14 (m, 1H) , 4.11-4.04 (m, 1H) , 1.53-1.46 (m, 2H) , 1.31-1.22 (m, 32H) , 1.13 (s, 6H) , 0.90 (t, J = 5.3 Hz, 3H) .

Example 3 Biochemical Assays

Assay 1: In vitro anti-viral activities against OC43

The in vitro anti-OC43 activity and cytotoxicity were evaluated using Huh7 cell as follows.

Anti-virial activity assay: The Huh7 cells were seeded in 96-well plates, in 100 μl per well of assay medium, at a density of 8000 cells/well and cultured at 37 ℃ and 5%CO2. After incubation for 24 hours, the test compound and reference compounds were diluted with assay medium and then added into the cells, 50 μl per well. Then 50 μl per well of assay medium diluted virus was added. The resulting cell culture are incubated for additional 7 days until virus infection in the virus control (cells infected with virus, without compound treatment) displays significant CPE (cytopathic effects) . The CPE are measured by CellTiter Glo following the manufacturer’s manual. The antiviral activity of compounds is calculated based on the protection of the virus-induced CPE at each concentration normalized by the virus control.

The %inhibition was calculated by the following equation: Inhibition (%) = (Raw data CPD -Average VC) / (Average CC -Average VC) *100.

Cytotoxicity assay: The cytotoxicity of compounds is assessed under the same method as in the anti-virial activity assay, but without the step of virus infection. The Huh7 cell viability is measured with Cell-Titer Glo following the manufacturer’s manual. The %Viability was calculated by the following equation: Viability (%) = (Raw data CPD -Average MC) / (Average CC -Average MC) *100.

Raw data CPD: values of the sample-treated wells

Average VC: average value of virus control

Average CC: average value of cell control (cells without virus infection or compound treatment)

Average MC: average value of medium control (medium only) wells.

EC50 and CC50 values was calculated using the GraphPad Prism software using the nonlinear regression model of log (inhibitor) vs. response --Variable slope (four parameters) .

Assay 2: In vitro anti-viral activities against 229E

The in vitro anti-229E activity and cytotoxicity were evaluated using MRC5 cell as follows.

Anti-virial activity assay: The MRC5 cells were seeded in 96-well plates, in 100 μl per well of assay medium, at a density of 20000 cells/well and cultured at 37 ℃ and 5%CO2. After incubation for 24 hours, the test compound and reference compounds were diluted with assay medium and then added into the cells, 50 μl per well. Then 50 μl per well of assay medium diluted virus was added. The resulting cell culture are incubated for additional 3 days until virus infection in the virus control (cells infected with virus, without compound treatment) displays significant CPE. The CPE are measured by CellTiter Glo following the manufacturer’s manual. The antiviral activity of compounds is calculated based on the protection of the virus-induced CPE at each concentration normalized by the virus control.

Cytotoxicity assay: The cytotoxicity of compounds is assessed under the same method as in the anti-virial activity assay, but without the step of virus infection. Cell viability is measured with Cell-Titer Glo following the manufacturer’s manual. The %Viability was calculated by the following equation: Viability (%) = (Raw data CPD -Average MC) / (Average CC -Average MC) *100.

Raw data CPD: values of the sample-treated wells

Average VC: average value of virus control

Average MC: average value of medium control (medium only) wells.

Assay 3: In vitro anti-viral activities against SARS-CoV-2

The in vitro anti-SARS-COV-2 activity and cytotoxicity were evaluated using Vero E6/HAE/A549 cells as follows.

Anti-virial activity assay: The Vero E6/HAE/A549 cells were seeded in 96-well plates, in 100 μl per well of assay medium, at a density of Vero E6/HAE/A549 cells/well and cultured at 37 ℃ and 5%CO2. After incubation for 24 hours, the test compound and reference compounds were diluted with assay medium and then added into the cells, 50 μl per well. Then 50 μl per well of assay medium diluted virus was added. The resulting cell culture are incubated for additional 3 days until virus infection in the virus control (cells infected with virus, without compound treatment) displays significant CPE. The CPE are measured by CellTiter Glo following the manufacturer’s manual. The antiviral activity of compounds is calculated based on the protection of the virus-induced CPE at each concentration normalized by the virus control.

Raw data CPD: values of the sample-treated wells

Average VC: average value of virus control

Average MC: average value of medium control (medium only) wells.

Assay 4: In vitro anti-viral activities against SARS-CoV-2 Replicon

The compounds were serially diluted in DMSO and added into 384-well plates 0.3 μl per well (8 doses, 3 fold, in duplicate wells) . The replicon RNA was generated in vitro transcript. Huh7 cells transfected with purified SARS-CoV-2 replicon RNAs were seeded 4000/well in 384 microplates containing serially diluted compounds, then cultured at 37℃ and 5%CO2 for 1 day. The final volume of the cell culture was 60μl per well, and the final concentrations of DMSO in the test plates was 0.5%.

Fluorescence intensity was determined using Acumen Cellista (TTP LabTech) , and the antiviral activity of compounds was calculated based on the inhibition of expression of GFP. Cell viability was measured with CellTiter Glo following the manufacturer’s manual.

The antiviral activity and viability of compounds were expressed as %Inhibition and %Viability, respectively, and calculated with the formulas below:
Inhibition (%) = (Raw data CPD -Average ZPE) / (Average HPE -Average ZPE)
*100
Viability (%) = (Raw data CPD -Average HPE) / (Average ZPE-Average HPE) *100

Raw data CPD: values of the sample-treated wells

Average ZPE: average value of virus control

Average HPE: average value of medium control (medium only) wells.

Assay 5: Determination of intracellular nucleoside triphosphates (NTP) of HepG2 and A549:

The human hepatocellular carcinoma cell line HepG2 and human lung adenocarcinoma cell line A549 were used to evaluate the drug cell uptake and intracellular nucleoside triphosphates. Cells were incubated in 6 well plate at 37℃ in a 5%CO2 -95%air atmosphere over allow the cells to attach. The cell density was 2×10⁶/well for hepG2 and 1x10⁶/well for A549. The incubation medium for HepG2 and A549 were Eagle's Minimum Essential Medium (EMEM) and F-12K, respectively, both containing 2%Fetal bovine serum (FBS) . After cell attached, the culture medium was removed and 2.5mL fresh medium which containing test compound at 1μM was add to each well for incubation at 37℃ in a 5%CO₂ -95%air atmosphere. At each specific time point (1, 2, 4, 6 h) , remove the culture medium and wash the cells with 3mL ice-cold PBS twice. After washing, the cells were digested from the plate wells with 600μL ice-cold 70%methanol. The cell lysates were then centrifuged at 4 ℃, 13000rpm for 5 min. An aliquot of (200uL) the supernatant was transferred to a clean 96 well plate for LC-MS/MS (Shimazu 20A and API 4000 Qtrap) analysis to determine the concentration of intracellular nucleoside triphosphates.

Assay 6: Stability in human liver S9 fraction:

Human or rat liver S9 were used for stability evaluation. In brief, compounds (1μM) were incubated in 0.2mL of 100mM potassium phosphate buffer (PPB, pH 7.4) containing S9 (2.0mg protein/mL) at 37℃. with time points taken at 0, 5, 15, 30, 45 and 60 minutes. After incubation at each time point, the reaction was stopped by adding 600μL ice cold acetonitrile. The samples were then centrifuged, and the supernatant was injected to UPLC-MS/MS for analysis. Half-life (t1/2) was determined from the disappearance rate constant (k) using the following equation: In vitro t1/2 = 0.693 /k. The disappearance rate constant (k) was determined as the gradient of the natural logarithm of the percentage remaining vs. incubation time curve by regression analysis, where the percentage remaining (%) was calculated as follows:

Tables 1-4 show the results of exemplary compounds of the present disclosure and the reference compounds (Remdesivir, Remdesivir monophosphate (Remdesivir-MP) , GS-441524 and ( (2R, 3S, 4R, 5R) -5- (4-aminopyrrolo [2, 1-f] [1, 2, 4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl (3- (hexadecyloxy) propyl) hydrogen phosphate (Reference Compound 1) ) for Assays 1-6. It shows the compounds of present disclosure have good inhibitory effect against various coronaviruses with low cytotoxicity, show higher stability in human liver and produce more triphosphorylated products in lung cells.

Assay 7: In vitro anti-viral activities against SARS-CoV-2 omicron BA. 5

The in vitro anti-SARS-COV-2 omicron BA. 5 activity and cytotoxicity of compounds were tested, in combination with 2 μM CP-100356, for in vitro antiviral activity against SARS-CoV-2, omicron BA. 5, in Vero E6 cells. Test media was MEM supplemented with 2%FBS and 50 μg/mL gentamicin.

CP-100356 was provided by the sponsor in powder form and was solubilized in DMSO to prepare a 10 mM solution. Test compounds and reference compounds were also received from sponsor in powder form and solubilized in DMSO to prepare 10 mM stock solutions. The latter compounds were serially diluted using eight half-log dilutions in test media containing CP-100356 (final on-plate 2 μM in all dilutions) so that the starting (high) concentration of test compounds was 1 μM. Each dilution was added to 5 wells of a 96-well plate with 80-100%confluent cells. Three wells of each dilution were infected with virus, and two wells remained uninfected as toxicity controls. Six wells were infected and untreated as virus controls, and six wells were uninfected and untreated as cell controls. Virus was prepared to achieve a MOI of 0.003. Remdesivir was tested in parallel as a positive control. Plates were incubated at 37±2℃, 5%CO2.

On day 3 post-infection, once untreated virus control wells reached maximum CPE, plates were stained with neutral red dye for approximately 2 hours (±15 minutes) . Supernatant dye was removed and wells rinsed with PBS, and the incorporated dye was extracted in 50: 50 Sorensen citrate buffer/ethanol for >30 minutes and the optical density was read on a spectrophotometer at 540 nm. Optical densities were converted to percent of cell controls and normalized to the virus control, then the concentration of test compound required to inhibit CPE by 50% (EC50) was calculated by regression analysis. The concentration of compound that would cause 50%cell death in the absence of virus was similarly calculated (CC50) . The selective index (SI) is the CC50 divided by EC50. Table 5 shows the in vitro anti-viral activities against SARS-CoV-2 Omicron BA. 5 for exemplary compounds.

Assay 8: Determination of intracellular nucleoside triphosphates in HAE

The human airway epithelial cells (HAECs, HAE) line was used to evaluate the drug cell uptaking and intracellular nucleoside triphosphates. Cells were incubated in a 6 well plate at 37℃ in a 5%CO₂ -95%air atmosphere overnight to allow the cells to attach. The cell density was 3x105/well for HAECs. The incubation medium for HAECs was special designed medium (CM-H016) containing 2%Fetal bovine serum (FBS) . After cell attached, the culture medium was removed and 2.5mL fresh medium which containing test compound at 5μM was add to each well for incubation at 37℃ in a 5%CO₂ -95%air atmosphere. At each specific time point (1, 2, 4, 6h) , remove the culture medium and wash the cells with 3mL ice-cold PBS twice. After washing, the cells were digested from the plate wells with 600μL ice-cold 70%methanol. The cell lysates were then centrifuged at 4℃, 13000rpm for 5min. An aliquot of (200μL) the supernatant was transferred to a clean 96 well plate for LC-MS/MS (Waters and API 5500) analysis to determine the concentration of intracellular nucleoside triphosphates. Table 6 shows concentration of intracellular nucleoside triphosphates for exemplary compounds.

Table 1 In vitro anti-viral activities against 229E for exemplary compounds

¹ For EC₅₀, A: <100 nM, B: 100-1000 nM, C: >1000 nM.

Table 2 In vitro anti-viral activities against SARS-CoV-2 Replicon for exemplary compounds

¹ For EC₅₀, A: <100 nM, B: 100-1000 nM, C: >1000 nM.

Table 3 Stability in human liver S9 fraction for exemplary compounds

¹ For t_1/2, A: >60 min, B: 20-60 min, C: <20 min.

Table 4 Intracellular nucleoside triphosphates for exemplary compounds

¹ For MetaPpp Concentration: A: >500nM, B: 200～500nM, C: <200nM.

Table 5 In vitro anti-viral activities against SARS-CoV-2 Omicron BA. 5 for exemplary compounds

¹ For EC₅₀, A: <50 nM, B: 50-500 nM, C: >500 nM.

Table 6 Intracellular nucleoside triphosphates for exemplary compounds

¹ For MetaPpp Concentration: A: >400nM, B: 200～400nM, C: <200nM.

The foregoing description is considered as illustrative only of the principles of the present disclosure. Further, since numerous modifications and changes will be readily apparent to those skilled in the art, it is not desired to limit the invention to the exact construction and process shown as described above. Accordingly, all suitable modifications and equivalents may be considered to fall within the scope of the invention as defined by the claims that follow.

Claims

A compound having Formula (I) :

or a pharmaceutically acceptable salt thereof,

wherein

Base is a naturally occurring or modified pyrimidine base or purine base;

R¹ is selected from the group consisting of hydrogen, halogen, hydroxyl, cyano, azido, alkyl, alkenyl, alkynyl, haloalkyl, -OR^a, -NO₂, -N (R^a) ₂, -C (O) N (R^a) ₂, -C (O) R^a, -OC (O) R^a, -C (O) OR^a, -S (O) R^a, -S (O) ₂R^a, -S (O) OR^a, -S (O) ₂ (OR^a) , and -S (O) ₂N (R^a) ₂;

each of R²¹, R²², R³¹, R³², and R⁴ is independently selected from the group consisting of hydrogen, deuterium, halogen, hydroxyl, cyano, azido, alkyl, alkenyl, alkynyl, haloalkyl, -OR^a, -NO₂, -N (R^a) ₂, -C (O) N (R^a) ₂, -C (O) R^a, -OC (O) R^a, -C (O) OR^a, -S (O) R^a, and -S (O) ₂R^a;

Q is CH₂, CHD or CD₂;

each M is independently selected from hydrogen, metal ion, -NH₄, or protonated organic amine;

L is selected from hydrogen or -C (R⁵) ₂-L¹-L²;

each R⁵ is selected from hydrogen, deuterium, or alkyl optionally substituted with one or more groups independently selected from halogen, hydroxyl, cyano, cycloalkyl, heterocyclyl, aryl or heteroaryl;

L¹ is selected from -OC (O) -*, -OC (O) O-*, -OC (O) N (R^a) -*, -N (R^a) C (O) -*, -N (R^a) C (O) O-*, orwherein *end of L¹ is connected to L²;

L² is selected from hydrogen, R⁶, wherein *end of L² is connected to L¹;

R⁶ is selected from C_9-26 alkyl, C_9-26 alkenyl, C_9-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, R^aOCH₂ (CH₂OCH₂) _nCH₂-, wherein the alkyl, alkenyl or alkynyl is optionally substituted with one or more R^b;

R⁷ is selected from C_6-26 alkyl, C_6-26 alkenyl, C_6-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-, each of which is optionally substituted with one or more R^b;

each of R⁸ and R⁹ is independently selected from hydrogen, C_1-26 alkyl, C_2-26 alkenyl, C_2-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-, each of which is optionally substituted with one or more R^b;

provided that when R⁸ is methyl and R⁹ is hydrogen, then R⁷ is selected from C_7-26 alkyl, C_7-26 alkenyl, C_7-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-;

each R^a is independently selected from hydrogen, halogen or alkyl;

each R^b is independently selected from halogen, hydroxyl, cyano, amino or alkyl;

m is 0, 1, 2, or 3; and

each n, p, or q is independently an integer between 0-20.
The compound according to claim 1, or a pharmaceutically acceptable salt thereof, having a Formula (Ia) :
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the base is selected from the group consisting of:
The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein the base is selected from the group consisting of:
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein L is hydrogen.
The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein m is 0, 1 or 3.
The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein M is hydrogen.
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein L is-C (R⁵) ₂-L¹-L².
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein both R⁵ are hydrogen or deuterium.
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein one R⁵ is hydrogen, and the other R⁵ is alkyl optionally substituted with aryl.
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein one R⁵ is hydrogen, and the other R⁵ is methyl, ethyl,
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein L¹ is selected from -OC (O) -*, -OC (O) O-*, -OC (O) NH-*, -OC (O) N (CH₃) -*, -NHC (O) -*, -N (CH₃) C (O) -*, -NHC (O) O-*, -N (CH₃) C (O) O-*, or
The compound of claim 12, or a pharmaceutically acceptable salt thereof, wherein L¹ is -OC (O) -*, -OC (O) O-*or
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein L² is R⁶.
The compound of claim 14, or a pharmaceutically acceptable salt thereof, wherein R⁶ is linear C_9-26 alkyl, linear C_9-26 alkenyl, linear C_9-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-.
The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R⁶ is linear C_14-22 alkyl.
The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R⁶ is linear C_14-22 alkenyl.
The compound of claim 17, or a pharmaceutically acceptable salt thereof, wherein R⁶ is linear C_14-22 alkenyl comprising 1-6 double bonds.
The compound of claim 18, or a pharmaceutically acceptable salt thereof, wherein each double bond is in Z configuration.
The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R⁶ is CH₃ (CH₂) _pO (CH₂) _q-.
The compound of claim 20, or a pharmaceutically acceptable salt thereof, wherein p is an integer from 3 to 20, and q is an integer from 1 to 6.
The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R⁶ is R^aOCH₂ (CH₂OCH₂) _nCH₂-.
The compound of claim 22, or a pharmaceutically acceptable salt thereof, wherein n is an integer from 1 to 6.
The compound of claim 22, or a pharmaceutically acceptable salt thereof, wherein R^a is hydrogen, methyl or ethyl.
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein L² is
The compound of claim 25, or a pharmaceutically acceptable salt thereof, wherein R⁷ is C_6-26 alkyl, C_6-26 alkenyl, C_6-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-.
The compound of claim 26, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_6-26 alkyl or linear C_6-26 alkenyl.
The compound of claim 27, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_13-21 alkyl or linear C_13-21 alkenyl.
The compound of claim 28, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_13-21 alkenyl comprising 1-6 double bonds.
The compound of claim 29, or a pharmaceutically acceptable salt thereof, wherein each double bond is in Z configuration.
The compound of claim 26, or a pharmaceutically acceptable salt thereof, wherein R⁷ is CH₃ (CH₂) _pO (CH₂) _q-.
The compound of claim 31, or a pharmaceutically acceptable salt thereof, wherein p is an integer from 3 to 20, and q is an integer from 1 to 6.
The compound of claim 26, or a pharmaceutically acceptable salt thereof, wherein R⁷ is R^aOCH₂ (CH₂OCH₂) _nCH₂-.
The compound of claim 33, or a pharmaceutically acceptable salt thereof, wherein n is an integer from 1 to 6.
The compound of claim 33, or a pharmaceutically acceptable salt thereof, wherein R^a is hydrogen, methyl or ethyl.
The compound of claim 25, or a pharmaceutically acceptable salt thereof, wherein R⁸ is C_1-26 alkyl or C_2-26 alkenyl, and R⁹ is hydrogen or C_1-26 alkyl.
The compound of claim 36, or a pharmaceutically acceptable salt thereof, wherein R⁸ is linear C_1-26 alkyl or linear C_8-26 alkenyl, and R⁹ is hydrogen or linear C_1-26 alkyl.
The compound of claim 25, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_6-26 alkyl, R⁸ is linear C_2-26 alkyl, and R⁹ is hydrogen.
The compound of claim 25, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_7-26 alkyl, R⁸ is methyl, and R⁹ is hydrogen or methyl.
The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein L² is
The compound of claim 40, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_6-26 alkyl, linear C_6-26 alkenyl, linear C_6-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-.
The compound of claim 41, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_6-26 alkyl or linear C_6-26 alkenyl.
The compound of claim 40, or a pharmaceutically acceptable salt thereof, wherein R⁸ is linear C_1-26 alkyl, linear C_2-26 alkenyl, linear C_2-26 alkynyl, CH₃ (CH₂) _pO (CH₂) _q-, or R^aOCH₂ (CH₂OCH₂) _nCH₂-, and R⁹ is hydrogen or C_1-26 alkyl.
The compound of claim 43, or a pharmaceutically acceptable salt thereof, wherein R⁸ is linear C_1-26 alkyl or linear C_2-26 alkenyl, and R⁹ is hydrogen or linear C_1-26 alkyl.
The compound of claim 40, or a pharmaceutically acceptable salt thereof, wherein R⁷ is linear C_6-26 alkyl, R⁸ is linear C_1-26 alkyl, and R⁹ is hydrogen.
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein R¹ is selected from hydrogen, hydroxyl, cyano, azido, alkyl, or haloalkyl.
The compound of claim 46, or a pharmaceutically acceptable salt thereof, wherein R¹ is cyano.
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein each of R²¹, R²², R³¹, R³², and R⁴ is independently selected from hydrogen, deuterium, halogen, hydroxyl, cyano, azido, alkyl, haloalkyl, -OC (O) R^a.
The compound of claim 48, or a pharmaceutically acceptable salt thereof, wherein each of R²¹, R²², R³¹, R³² is selected from hydrogen, deuterium, halogen, hydroxyl, alkyl or -OC (O) R^a.
The compound of claim 49, or a pharmaceutically acceptable salt thereof, wherein R^a is alkyl.
The compound of claim 50, or a pharmaceutically acceptable salt thereof, wherein R^a is methyl.
The compound of claim 48, or a pharmaceutically acceptable salt thereof, wherein R⁴ is selected from hydrogen, deuterium, halogen, cyano, azido, or haloalkyl.
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of:
The compound of any one of preceding claims, or a pharmaceutically acceptable salt thereof, wherein the base is is
The compound of claim 54, or a pharmaceutically acceptable salt thereof, wherein the base is
The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
A pharmaceutical composition comprising the compound of any one of claims 1-56 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
The pharmaceutical composition of claim 57, which is formulated for oral administration, injection administration, nasal administration, or pulmonary administration.
A method for treating a viral infection in a subject in need thereof, comprising administering an effective amount of a compound of any one of claims 1-56 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of any one of claims 57-58 to the subject.
The method of claim 59, wherein the viral infection is an RNA virus.
The method of claim 60, wherein the RNA virus is a single stranded or a double stranded RNA virus.
The method of claim 61, wherein the viral infection is from a virus selected from the group consisting of Retroviridae virus, Lentiviridae virus, Coronaviridae virus, Picornaviridae virus, Caliciviridae virus, Flaviviridae virus, Togaviridae virus, Bomaviridae, Filoviridae, Paramyxoviridae, Pneumoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Orthomyxoviridae, and Deltavirus.
The method of claim 62, wherein the viral infection is from a virus selected from the group consisting of Lymphocytic choriomeningitis virus, Coronavirus, HIV, SARS, Poliovirus, Rhinovirus, Hepatitis A, Norwalk virus, Yellow fever virus, West Nile virus, Hepatitis C virus, Dengue fever virus, Zika virus, Rubella virus, Ross River virus, Sindbis virus, Chikungunya virus, Boma disease virus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipah virus, Hendra virus, Newcastle disease virus, Human respiratory syncytial virus, Rabies virus, Lassa virus, Hantavirus, Crimean-Congo hemorrhagic fever virus, Influenza and Hepatitis D virus.
The method of claim 61, wherein the viral infection is a coronavirus infection.
The method of claim 64, wherein the coronavirus is selected from the group consisting of 229E alpha coronavirus, NL63 alpha coronavirus, OC43 beta coronavirus, HKU1 beta coronavirus, Middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV) , severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) , and SARS-CoV-2 (COVID-19) .
The method of claim 65, wherein the coronavirus is SARS-CoV-2.
A method for inhibiting RNA polymerase in a subject in need thereof, comprising administering an effective amount of a compound of any one of claims 1-56 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of any one of claims 57-58 to the subject.
Use of a compound of any one of claims 1-56 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of any one of claims 57-58, in the manufacture of a medicament for treating a viral infection.
A compound of any one of claims 1-56 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of any one of claims 57-58, for treating a viral infection.