Uridine phosphorylase (UPASE) inhibitors for treatment of liver diseases

Abstract

The present invention relates to uridine phosphorylase (UPASE) inhibitors for use in the treatment of liver diseases. The present invention provides methods of treating a liver condition (e.g., NAFLD, NASH, and / or DILI) in a subject. Aspects of the methods include administering to the subject an effective amount of a UPase inhibitor, optionally in combination with a uridine active agent (e.g., uridine (UR), a UR prodrug, or a UR mimetic), such as a supplemental uridine, to treat the liver condition in the subject. The invention also provides a composition for carrying out the subject method.

Description

[0001] This application is a divisional application. The application date of the original application is June 18, 2020, the application number is 202080058787.0 (PCT / US2020 / 038353), and the name of the invention is “Uridine phosphorylase (UPASE) inhibitors for the treatment of liver diseases”.

[0002] Cross-references to related patent applications

[0003] This patent application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 62 / 864,695, filed on June 21, 2019; the contents of which are incorporated herein by reference.

[0004] introduce

[0005] The prevalence of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), a more severe form of NAFLD, is increasing rapidly worldwide. 1 NAFLD is characterized by accumulation of fat in the liver (steatosis), occasional fibrosis, and hepatocyte swelling due to accumulation of fat (triglyceride) droplets in the liver without secondary causes of liver fat accumulation (eg, alcohol, infection, drugs). 2 In addition to hepatocyte swelling and fat accumulation such as those found in NALFD, NASH is characterized by lobular inflammation, fibrosis, and hepatocyte degeneration. 2 In NASH, fibrosis is often followed by cirrhosis and end-stage liver disease, which are often fatal without liver transplantation. 3 In addition to end-stage liver disease, individuals with NASH also often develop liver cancer (hepatocellular carcinoma, HCC) as a result of the condition. 4,5

[0006] The increasing prevalence of NALFD and NASH reflects the increasing prevalence of obesity and type 2 diabetes (T2D) in society and the hepatic manifestations of the resulting altered metabolic state. 6 Exposure of hepatocytes to high concentrations of lipids and carbohydrates (termed lipotoxicity and glucotoxicity, respectively) is responsible for much of the hepatocyte injury observed in NAFLD / NASH. 7 Metabolic syndrome, defined as a cluster of conditions including obesity, insulin resistance, hyperglycemia, dyslipidemia, and hypertension, is a major risk factor that predisposes individuals to NAFLD and NASH. 8 About 30% of North Americans have NALFD, and about 4% of North Americans have NASH. 9Genetic, demographic, and ethnic factors may also play a role in the pathogenesis of NAFLD,10 for example:

[0007] NAFLD has been associated with multiple genetic variants, including PNPLA-3, TM6SF2, and FDFT1.

[0008] NAFLD is more common in older people and in men.

[0009] In the United States, Hispanics have the highest prevalence of NAFLD, followed by Caucasians and then African Americans.

[0010] There are currently no drugs approved by the U.S. Food and Drug Administration (FDA) to treat NASH patients. Current recommended treatments for the disease include weight loss and dietary modifications, such as reducing fat and glucose consumption. 11 Given that the only known “cure” for advanced NASH and / or HCC is liver transplantation, it is inevitable that NASH will soon surpass hepatitis as the single leading reason for liver transplantation. 9

[0011] Lipid droplets (LDs), also known as liposomes, oil bodies, or fat bodies, are dynamic organelles that store neutral lipids during periods of energy surplus and serve as energy reservoirs during periods of energy deficit. 12 Many common metabolic diseases, such as metabolic syndrome and obesity, often lead to elevated liver lipids and increased LD, also known as hepatic steatosis / NAFLD. LD, especially LD-associated proteins, are closely related to the pathophysiology of fatty liver disease. 13 Under normal physiological conditions, liver LD is small and limited in number. Larger amounts of LD are associated with hepatotoxicity in animals or humans following administration of drugs such as tamoxifen (TAM). 14,15,16,17 Cyclosporine, 18 Valproic acid 19,20 ,tetracycline 21 , clofibrate 22 Olanzapine 23 and statins (such as simvastatin) 24 etc. LDs appear to have an intimate relationship with mitochondria, including physical contacts and protein shuttling.

[0012] 25 LDs also appear to have a similarly close relationship with the cell nucleus. 26 The content of LD seems to determine its potential toxicity - it contains unsaturated fatty acids (e.g. arachidonic acid, a precursor of many inflammatory mediators) 27The toxicity of LD is stronger than that of LD containing saturated fatty acids. 28 LDs are coated by proteins from the perilipin family, some of which are involved in the regulation of lipid metabolism. 29

[0013] The major public health problem related to the liver is drug-induced hepatitis (DILI), which affects individuals taking medications or nutraceuticals. DILI has been associated with more than 1,000 medications. 30 Although patients with milder forms of DILI can expect to recover completely, associated symptoms (e.g., fatigue, itching, nausea) can be debilitating and prolong recovery, and approximately 20% of patients have biochemical evidence of persistent liver damage six months after diagnosis. 31 Cirrhosis and long-term liver-related morbidity and mortality occur in approximately 3% of cases. 32 There is currently no test that doctors can use to definitively diagnose DILI. 33 DILI, or even suspected DILI, may lead to alternative treatment options, resulting in exposure to new adverse drug event risks and potentially suboptimal treatment of the underlying disease. DILI is also a common reason for termination of clinical drug development programs. 34 DILI causes a range of pathologies, including elevated serum aminotransferase levels detected on routine biochemical laboratory tests that resolve after elimination of the liver-damaging chemical agent, acute liver failure, defined as new-onset, sudden, and life-threatening liver dysfunction with coagulopathy and hepatic encephalopathy within 26 weeks of onset, and more. 35 Acute liver failure is a devastating disease as it primarily affects young, healthy individuals and results in the death of approximately 30% of patients who undergo invasive treatment, including liver transplantation.

[0014] DILI is divided into two types: "intrinsic" and "idiosyncratic".

[0015] Drugs that induce liver injury in a predictable, dose-dependent manner in preclinical models and humans are claimed to cause intrinsic DILI. Acetaminophen is the most common cause of intrinsic DILI in the United States. Few other drugs on the market are known to cause life-threatening intrinsic DILI, as this tendency is usually identified in preclinical or early clinical studies. Such drugs are typically not further developed, are used at doses that are optimal for efficacy but are not expected to cause liver injury, or are administered in controlled or desperate circumstances, such as chemotherapy. There are no available treatment options for intrinsic DILI other than N-acetylcysteine ​​for the treatment of acetaminophen overdose, and N-acetylcysteine ​​is indicated for limited clinical settings.

[0016] Idiosyncratic DILI is the most problematic form of DILI. It is rare in treated patients and usually occurs several months after treatment with the liver-damaging drug. Idiosyncratic DILI caused by a new drug is usually not discovered until the drug is widely used.

[0017]

[0018] Idiosyncratic DILI with a latent period may reflect an immune attack on the liver. Consistent with this view, if patients with DILI resume the use of liver-damaging drugs, idiosyncratic liver injury will recur rapidly after full recovery. The prolonged latent period can be attributed to the time required for antigen-specific lymphocytes to activate and proliferate to sufficient numbers to mediate DILI. The first step in initiating idiosyncratic DILI may be the formation of hepatocyte stress-induced neoantigens.

[0019] Both intrinsic and idiopathic liver injury appear to follow a similar process. Proposed mechanisms include mitochondrial dysfunction, oxidative stress, and altered bile acid homeostasis. Mitochondria produce the ATP required to maintain all vital cellular functions. Drugs that induce DILI can inhibit mitochondrial function, resulting in reduced ATP levels, decreased cellular function, and ultimately cell death. 36 Oxidative stress is caused by ROS, which are byproducts of normal metabolism and play a role in cell signaling and homeostasis. However, some drugs that induce DILI can increase ROS accumulation through multiple mechanisms. 37 When established processes that regulate cellular ROS levels are exceeded, oxidative stress can lead to damage to key cellular components and ultimately cell death. Finally, a major function of the liver is to transport bile salts from the blood into the bile. Drugs that induce DILI can disrupt this process in a variety of ways, most importantly by reducing hepatic bile acid excretion through inhibition of bile salt exporters. 38 This allows the accumulation of toxic bile acids within the cell, which can lead to hepatocyte death. In summary, DILI is classified into two different types - intrinsic or idiosyncratic, which share similar pathology and pathogenesis but may have multiple initiating and promoting mechanisms.

[0020] The association between NAFLD / NASH and DILI is not fully understood. 39,40,41,42,43,44However, (1) DILI is a risk factor for patients with NAFLD / NASH for many drugs, (2) DILI manifests as lesions similar to those in NAFLD / NASH, (3) the pathophysiology of DILI and NASH overlaps, (4) certain drugs induce hepatic steatosis (DIS) and / or steatohepatitis (DISH) by triggering pathological events similar to those that occur in the development and progression of NAFLD / NASH, (5) DILI affects the development of NAFLD / NASH or accelerates the progression of NAFLD / NASH, and (6) NAFLD / NASH influences susceptibility to DILI and the prognosis of DILI.

[0021] The deleterious effects of DILI may not only mimic the physiological insults in the normal liver that drive the development and / or progression of NAFLD, but may also exacerbate similar pre-existing alterations in fatty liver. Relative to NAFLD / NASH, DILI:

[0022] Aggravating factors (e.g. obesity, diabetes)

[0023] Enhances steatosis factors (e.g., increases hepatic lipid synthesis / uptake)

[0024]

[0025] Increased inflammatory factors (e.g., accumulation of lipotoxic fatty acids and oxidative stress)

[0026] Activation of profibrotic factors (e.g., increased collagen deposition)

[0027] Altered drug-metabolism system in the context of NAFLD

[0028] Based on the above, agents that can alleviate NAFLD / NASH can also be used to treat DILI, and vice versa.

[0029] NAFLD / NASH has become a significant and potentially fatal epidemic for which there are no approved treatment options in countries with increasing prevalence of obesity and related metabolic disorders. DILI is another serious liver disorder that appears to have a similar pathophysiology to NAFLD / NASH. Summary of the invention

[0030] The present invention provides a method for treating a subject's liver disorder (e.g., NAFLD, NASH and / or DILI). Aspects of the method include administering an effective amount of a UPase inhibitor to the subject, optionally in combination with a uridine active agent (e.g., uridine (UR), UR prodrug or UR mimetic), such as supplementing uridine, to treat the subject's liver disorder. The present invention also provides a composition for implementing the subject's method. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 A regression analysis of plasma UR concentrations measured after continuous infusion of different amounts of compound 1 (TK-112690) into mice and plasma compound 1 concentrations is provided. The R2 of the straight line is 0.95, and the slope and intercept values ​​of the straight line are 0.010 and 0.051, respectively. Compound 1 was observed to increase plasma UR in a linear manner.

[0032] Figure 2 The weights of mice measured during the experimental phase of the MCD study are provided. Six mice per experimental group were studied. Significant weight loss occurred in each group. No changes in body weight between groups were observed. Data are expressed as mean + / - SEM.

[0033] Figure 3 Serum HDL cholesterol levels measured at the end of the experimental phase of the MCD study are provided. Six mice per experimental group were studied. HDL cholesterol levels in mice receiving UR + Compound 1 (TK-112690) were significantly reduced (p<0.05) compared to control animals receiving vehicle administration. Data are expressed as mean + / - SEM.

[0034] Figure 4 Provided are (A) representative H&E images of liver sections from each experimental group and (B) fibrosis scores for the images shown in (A). The most significant effect was in the group of mice that received UR + compound 1 (TK-112690). Compared with the control group that received vehicle administration, liver fibrosis in this group was significantly reduced (p < 0.001) (six animals per experimental group). Data are expressed as mean + / - SEM.

[0035] definition

[0036] In describing the compounds, pharmaceutical compositions containing such compounds, methods of use of such compounds and compositions, and biological and pharmacological contexts involving the use of the compounds, the following terms have the following meanings unless otherwise indicated. It should also be understood that any moiety defined below may be substituted with a variety of substituents, and that the respective definitions are intended to include such substituted moieties within their respective scopes.

[0037] "Acyl" refers to the group -C(O)R, where R is hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroalkyl, or heteroaryl as defined herein. Representative examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl, and the like.

[0038] "Acylamino" refers to the group -NR'C(O)R, where R' is hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, heteroaralkyl, and R is hydrogen, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, or heteroaralkyl as defined herein. Representative examples include, but are not limited to, formylamino, acetylamino, cyclohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino, benzylcarbonylamino, and the like.

[0039] "Acyloxy" refers to the group -OC(O)H, -OC(O)-alkyl, -OC(O)-aryl or -OC(O)-cycloalkyl.

[0040] "Aliphatic" refers to hydrocarbon-based organic compounds or groups characterized by a straight chain, branched chain, or cyclic arrangement of the constituent carbon atoms, and the absence of aromatic unsaturation. Aliphatics include, but are not limited to, alkyl, alkylene, alkenyl, alkenylene, alkynyl, and alkynylene. Aliphatic groups typically have from 1 or 2 to 6 or 12 carbon atoms.

[0041] "Alkenyl" refers to a monovalent ethylenically unsaturated hydrocarbon radical having up to about 11 carbon atoms, particularly 2 to 8 carbon atoms, more particularly 2 to 6 carbon atoms, which may be straight chain or branched and has at least 1, particularly 1 to 2 sites of ethylenic unsaturation. Specific alkenyl groups include vinyl (-CH=CH2), n-propenyl (-CH2CH=CH2), isopropenyl (-C(CH3)=CH2), vinyl and substituted vinyl, and the like.

[0042] "Alkoxy" refers to the group -O-alkyl. For example, specific alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0043] "Alkoxycarbonyl" means the group -C(O)-alkoxy where alkoxy is as defined herein.

[0044] "Alkoxycarbonylamino" refers to the group -NRC(O)OR' where R is hydrogen, alkyl, aryl or cycloalkyl and R' is alkyl or cycloalkyl.

[0045] "Alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, especially having up to about 12 or 18 carbon atoms, more especially having 1 to 8 carbon atoms as a lower alkyl group, and more especially having 1 to 6 carbon atoms. The hydrocarbon chain can be straight or branched. This term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl, n-octyl, tert-octyl, etc. The term "alkyl" also includes "cycloalkyl" as defined herein. The structures of some exemplary alkyl groups are shown in Table 1 below.

[0046]

[0047] "Alkylene" refers to a divalent saturated aliphatic hydrocarbon radical having, in particular, up to about 12 or 18 carbon atoms, more in particular 1 to 6 carbon atoms, which may be straight or branched. This term is exemplified by radicals such as methylene (-CH2-), ethylene (-CH2CH2-), propylene isomers (e.g., -CH2CH2CH2- and -CH(CH3)CH2-).

[0048] "Alkynyl" refers to an acetylenic unsaturated hydrocarbon group, especially having up to about 12 or 18 carbon atoms, more especially 2 to 6 carbon atoms, which group can be straight chain or branched and has at least 1, more especially 1 to 2 alkynyl unsaturation sites. Non-limiting examples of specific alkynyl groups include acetylenic groups, ethynyl (-C≡CH), propargyl (-CH2C≡CH), etc.

[0049] "Amino" refers to the group -NH2.

[0050] "Amino acid" refers to any naturally occurring amino acid (e.g., Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Hyl, Hyp, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val) in D, L, or DL ​​form. The side chains of naturally occurring amino acids are well known in the art and include, for example, hydrogen (e.g., in glycine), alkyl (e.g., in alanine, valine, leucine, isoleucine, proline), substituted alkyl (e.g., in threonine, serine, methionine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, arginine, and lysine), alkaryl (e.g., in phenylalanine and tryptophan), substituted arylalkyl (e.g., in tyrosine), and heteroarylalkyl (e.g., in histidine).

[0051] "Aminocarbonyl" refers to the group -C(O)NRR where each R is independently hydrogen, alkyl, aryl or cycloalkyl, or where the R groups are linked to form an alkylene group.

[0052] "Aminocarbonylamino" refers to the group -NRC(O)NRR where each R is independently hydrogen, alkyl, aryl or cycloalkyl, or where two R groups are linked to form an alkylene group.

[0053] "Aminocarbonyloxy" refers to the group -OC(O)NRR where each R is independently hydrogen, alkyl, aryl or cycloalkyl, or where the R groups are linked to form an alkylene group.

[0054] "Amino group-containing sugar group" refers to a sugar group having an amino group substituent. Representative amino group-containing sugar groups include L-vancomosamine, 3-demethyl-vancomosamine, 3-epi-vancomosamine, 4-epi-vancomosamine, acosamine, actinosamine, daunosamine, 3-epi-daunosamine, ristosamine, N-methyl-D-glucosamine, and the like. "

[0055] "ARMD" refers to an eye disease called age-related macular degeneration.

[0056] "Aralkyl" or "arylalkyl" refers to an alkyl group as defined above substituted with one or more aryl groups as defined above.

[0057] "Aryl" refers to a monovalent aromatic hydrocarbon group derived from a single carbon atom in a parent aromatic ring system after removing a hydrogen atom from the parent aromatic ring system. Typical aryl groups include, but are not limited to, groups derived from the following: anthracene, acenaphthylene, anthracene, azulene, benzene, lettuce, coronene, fluoranthene, fluorene, hexacene, hexylene, hexalene, asymmetric indacene, symmetric indacene, indane, indene, naphthalene, octacene, octalene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenanthene, phenanthrenes, phenanthrenes, pyrenes, pyranthene, rubine, triphenylene, trinaphthalene, etc. In particular, aryl groups contain 6 to 14 carbon atoms. The structures of some exemplary aryl groups are shown in Table 2 below.

[0058]

[0059]

[0060] "Aryloxy" means an -O-aryl group where "aryl" is as defined herein.

[0061] An "autoimmune disease" or "autoimmune condition" refers to a disease that occurs when the body's tissues are attacked by its own immune system. Examples of autoimmune diseases or conditions include multiple sclerosis, ankylosing spondylitis, Crohn's disease, arthritis, psoriasis, Behcet's disease, and psoriatic arthritis.

[0062] "Azide" refers to the group -N3.

[0063] "Carbohydrate" refers to a monosaccharide, disaccharide, trisaccharide or polysaccharide, wherein the molecular weight of the polysaccharide may be at most about 20,000, for example, hydroxypropyl-methylcellulose or chitosan. "Carbohydrate" also encompasses any atom of the sugar moiety such as an oxidized, reduced or substituted sugar monovalent group covalently linked to an anhydropyrimidine (e.g., anhydrothymidine or anhydrouridine) or its derivatives through an aglycone carbon atom. "Monosaccharide, disaccharide, trisaccharide or polysaccharide" may also include a glycosyl group containing an amino group. Representative "carbohydrates" include, for example, hexoses such as D-glucose, D-mannose, D-xylose, D-galactose, vancomycin, 3-demethyl-vancomycin, 3-epi-vancomycin, 4-epi-vancomycin, acosamine, actinosamine, daunosamine, 3-epi-daunosamine, ristosamine, D-glucosamine, N-methyl-D-glucosamine, D-glucuronic acid, N-acetyl-D-glucosamine, N-acetyl-D-galactose. Amine, sialic acid, iduronic acid, L-fructose, etc.; pentoses such as D-ribose or D-arabinose; ketoses such as D-ribulose or D-fructose; disaccharides such as 2-O-(α-L-vancosamino)-β-D-pyranose glucose-, 2-O-(3-demethyl-α-L-vancosamino)-β-D-pyranose glucose, sucrose, lactose or maltose; derivatives such as acetals; amines; acylated, sulfated and phosphorylated sugars; oligosaccharides having 2 to 10 sugar units. The sugar may be in its open-ring, r-pyranose or furanose form.

[0064] "Carboxy" refers to the group -C(O)OH.

[0065] "Cyano" refers to the radical -CN.

[0066] "Cycloalkenyl" refers to a cyclic hydrocarbon radical having 3 to 10 carbon atoms, having a single ring or multiple condensed rings (including fused and bridged ring systems), and having at least one, especially 1 to 2, sites of olefinic unsaturation. Such cycloalkenyl radicals include, for example, single ring structures such as cyclohexenyl, cyclopentenyl, cyclopropenyl, and the like.

[0067] "Cycloalkyl" refers to a cycloalkyl group having 3 to about 10 carbon atoms and having a single ring or multiple condensed rings (including fused and bridged ring systems), which may be optionally substituted with 1 to 3 alkyl groups. Such cycloalkyl groups include, for example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc.; and polycyclic structures such as adamantyl, etc.

[0068] "DILI" refers to drug-induced liver injury.

[0069] "DIS" refers to drug-induced steatosis.

[0070] "DISH" stands for drug-induced steatohepatitis.

[0071] “DR” refers to the eye condition – diabetic retinopathy.

[0072] "FU" refers to 5-fluorouracil.

[0073] "Heterocycloalkyl" refers to stable non-aromatic heterocyclic rings and fused rings containing one or more heteroatoms independently selected from N, O and S. The fused heterocyclic ring system may include carbocyclic rings and need only include one heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, piperazinyl, homopiperazinyl, piperidinyl and morpholinyl. The structures of some exemplary heterocyclic groups are shown in Table 3.

[0074]

[0075] "Halo" or "halogen" refers to fluoro, chloro, bromo and iodo. A halo group can be fluoro or chloro.

[0076] "HCC" refers to hepatocellular carcinoma.

[0077] "HDL" refers to high-density lipoprotein.

[0078] When used to describe a compound or a group present on a compound, "hetero" means that one or more carbon atoms in the compound or group are replaced by nitrogen, oxygen or sulfur heteroatoms. Hetero can be used for any of the above-mentioned hydrocarbyl groups having 1 to 5, especially 1 to 3 heteroatoms, such as alkyl, such as heteroalkyl; cycloalkyl, such as heterocycloalkyl; aryl, such as heteroaryl; cycloalkenyl, such as heterocycloalkenyl; cycloheteroalkenyl, such as heterocycloheteroalkenyl, etc. A heteroatom is any atom other than carbon or hydrogen, and is typically, but not limited to, nitrogen, oxygen, sulfur, phosphorus, boron, chlorine, bromine or iodine. Unsubstituted heteroatoms refer to side heteroatoms, such as amines, hydroxyls and thiols. Substituted heteroatoms refer to heteroatoms other than side heteroatoms.

[0079] "Heteroaryl" refers to a monovalent heteroaryl radical derived by removing a hydrogen atom from a single atom of a parent heteroaromatic ring system. Typical heteroaryl radicals include, but are not limited to, radicals derived from acridine, arsinole, carbazole, β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, The heteroaryl groups may be 5-20 yuan heteroaryl or 5-10 yuan heteroaryl. Specific heteroaryl groups are groups derived from thiophene, pyrrole, benzothiophene, benzofuran, indole, pyridine, quinoline, imidazole, oxazole and pyrazine.

[0080] "Hydroxy" refers to the radical -OH.

[0081] "KO" refers to gene knockout as used in the phrase "knockout animal".

[0082] "MCD" refers to the methionine-choline deficient diet.

[0083] "NAFLD" refers to non-alcoholic fatty liver disease.

[0084] "NASH" stands for nonalcoholic steatohepatitis.

[0085] "Nitro" refers to the radical -NO2.

[0086] "Peptide" refers to a polyamino acid containing up to 2, 5, 10 or about 100 amino acid residues.

[0087] "Polypeptide" refers to a polyamino acid containing about 100 amino acid units to about 1,000 amino acid units, about 100 amino acid units to about 750 amino acid units, or about 100 amino acid units to about 500 amino acid units.

[0088] “ROP refers to an eye condition that occurs in infants – retinopathy of prematurity.

[0089] "SEM" refers to standard error of the mean.

[0090] "Side effect" refers to an undesired adverse consequence of a drug administration.

[0091] A "stereoisomer" with respect to a given compound is well known in the art and refers to another compound having the same molecular formula in which the atoms making up the other compound are oriented in space in a different manner, but in which the atoms in the other compound are identical to the atoms in the given compound with respect to the atoms to which they are attached (e.g., enantiomers, diastereomers, or geometric isomers). For example, Morrison and Boyd, Organic Chemistry, 1983, 4th edition, Allyn and Bacon, Inc., Boston, MA, p. 123.

[0092] "Substituted" refers to a group in which one or more hydrogen atoms are each independently replaced with the same or different substituents. A "substituted" group refers in particular to a group having one or more substituents, for example, 1 to 5 substituents, and in particular 1 to 3 substituents, selected from the group consisting of acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, aralkyl, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxy, imido, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, sulfhydryl, alkylthio, (substituted alkyl)thio, arylthio, (substituted aryl)thio, alkyl-S(O)-, aryl-S(O)-, alkyl-S(O)2- and aryl-S(O)2. Typical substituents include, but are not limited to -X, -R 8 (provided that R8 is not hydrogen), -O-, =O, -OR 8 、-SR 8 , -S - , =S, -NR 8 R 9 , =NR 8 , -CX3, -CF3, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)2O - 、-S(O)2OH、-S(O)2R 8 、-OS(O2)O - 、-OS(O)2R 8 、-P(O)(O-)2、-P(O)(OR 8 )(O - )、-OP(O)(OR 8 )(OR 9 )、-C(O)R 8 、-C(S)R 8 、-C(O)OR 8 、-C(O)NR 8 R 9 、-C(O)O - 、-C(S)OR 8 、-NR 10 C(O)NR 8 R 9 、-NR 10 C(S)NR 8 R 9 、-NR 11 C(NR 10 )NR 8 R 9and -C(NR 10 )NR 8 R 9 , wherein X is independently halogen.

[0093] "Substituted amino" includes the groups cited in the definition of "substituted" herein, and particularly refers to the group -N(R)2, wherein each R is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, cycloalkyl, substituted cycloalkyl, and wherein two R groups are linked to form an alkylene group.

[0094] “T2D” refers to type 2 diabetes.

[0095] "TG" means genetically modified.

[0096] "Thioalkoxy" refers to the group -S-alkyl.

[0097] "Thioaryloxy" refers to an -S-aryl group.

[0098] "Thioketo" refers to the group =S.

[0099] "Sulhydryl" refers to the radical -SH.

[0100] "UR" refers to uridine.

[0101] "UPase (uridine phosphorylase)" in enzymology refers to a phosphorylase (EC 2.4.2.3) that catalyzes the chemical reaction: uridine + phosphate → uracil + α-D-ribose 1-phosphate. The two substrates of this enzyme are uridine and phosphate, while its two products are uracil and α-D-ribose 1-phosphate. This enzyme belongs to the family of glycosyltransferases, especially the pentosyltransferase family. The systematic name for this class of enzymes is uridine:phosphate α-D-ribosyltransferase. Other commonly used names include pyrimidine phosphorylase, UrdPase, UPH, and UPase. This enzyme is involved in pyrimidine metabolism.

[0102] "Uridine supplement" refers to a preparation containing UR or a preparation containing a UR precursor (e.g., monophosphate UR or acetylated UR) that is converted to UR in vivo. The preparation can be a solution, capsule, tablet, or cream. The product can be administered orally, intraperitoneally, subcutaneously, or intravenously. The UR supplement can be administered as part of a more complex mixture (e.g., a nutritional supplement).

[0103] ip, po and sc are intraperitoneal, oral or subcutaneous administration, respectively. H&E is hematoxylin-eosin, a dye used for tissue staining. SD is standard deviation. SE is standard error. PBS is phosphate-buffered saline. qd. and bid are once a day and twice a day, respectively.

[0104] Those skilled in the art will recognize that the maximum number of heteroatoms in a stable chemically feasible heterocyclic ring, whether aromatic or non-aromatic, is determined by the size of the ring, the degree of unsaturation of the heteroatoms, and the chemical valence. Generally, a heterocyclic ring may have from one to four heteroatoms as long as the heteroaromatic ring is chemically feasible and stable. Implementation

[0106] The present invention provides a method for treating a subject's liver disorder (e.g., NAFLD, NASH and / or DILI). Aspects of the method include administering an effective amount of a UPase inhibitor to the subject, optionally in combination with a uridine active agent (e.g., uridine (UR), UR prodrug or UR mimetic), such as supplementing uridine, to treat the subject's liver disorder. The present invention also provides a composition for implementing the subject's method.

[0107] Before describing the present invention in more detail, it should be understood that the present invention is not limited to the specific embodiments described, because there will be differences in actual implementation. It should also be understood that the terms used herein are only used to describe specific embodiments, and are not intended to limit the inventive concept, and the scope of the present invention will only be limited by the appended claims.

[0108] Where a numerical range is provided, it is to be understood that each intermediate value between the upper and lower limits of the range and any other specified value or intermediate value within the range are included within the scope of the present invention. Unless the context clearly specifies otherwise, each intermediate value should be as low as one-tenth of the lower limit unit. The upper and lower limits of these smaller ranges may be independently included within the smaller ranges, and are also included within the present invention, subject to the requirements of any specifically excluded limits within the range. Where the range includes one or two limits, the range excluding any one or two of those included limits is also included within the present invention.

[0109] Certain ranges set forth herein are preceded by the term "about." The term "about" is used herein for the purpose of providing literal support for the exact number that follows, as well as for numbers that are close to or approximately the number that follows the term. In determining whether a number is close to or approximately a specifically recited number, a close or approximately unrecited number may be a number that is substantially equivalent to the specifically recited number in the context in which it appears.

[0110] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those of ordinary skill in the art to which the invention belongs. Although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, representative exemplary methods and materials are described below.

[0111] All publications and patents cited in this specification are incorporated herein by reference as if each individual publication or patent was specifically and individually indicated to be incorporated by reference, and the purpose of incorporating by reference into this document is to disclose and describe the methods and / or materials related to the cited publications. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by reason of prior invention. In addition, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0112] It is noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is also noted that claims may be drafted to exclude any optional elements. Thus, this statement is intended to serve as antecedent basis for use of special terminology such as "solely," "only," etc. in connection with reciting claim elements or use of a "negative" limitation.

[0113] After reading the present invention, the following will be apparent to those skilled in the art that each individual embodiment described and listed herein has layered components and features that can be quickly decomposed or combined with the features of any of the other several embodiments without departing from the scope and spirit of the present invention. Any stated method can be implemented in the order of events stated or in any other order that is logically possible.

[0114] Although the apparatus and method have been or will be described for grammatical fluency and functional description, it should be clearly understood that unless otherwise expressly provided in Chapter 35 of the United States Code, the claims shall not be construed as necessarily being limited to "means" or "steps" but shall be consistent with the meaning and full scope of equivalents in accordance with the judicial doctrine of equivalents, and when the claims are expressly drafted in accordance with the provisions of Chapter 35 of the United States Code, the claims shall be fully consistent with the statutory equivalents in Chapter 35 of the United States Code.

[0115] In further describing the present invention, the subject methods are first described in more detail, then various compositions (eg, formulations and kits) that can be used in the subject methods are discussed, and various representative applications using the subject methods and compositions are discussed.

[0116] method

[0117] As described above, the present invention provides a method for treating, including prophylactic treatment (e.g., preventing occurrence) of a subject's liver condition. Aspects of the method include administering an UPase inhibitor to the subject alone or in combination with a uridine (UR) active agent (e.g., UR, UR prodrug, or UR mimetic) to treat the subject's liver condition. When the UPase inhibitor is co-administered with the UR active agent, i.e., administered simultaneously, the UPase inhibitor can be administered synchronously with the UPase inhibitor. Alternatively, the UPase inhibitor and the UR active agent can be administered sequentially, for example, the UPase inhibitor is administered before the UR active agent or the UPase inhibitor is administered after the UR active agent. In such embodiments, the UPase inhibitor and the UR active agent can be administered simultaneously, for example, as two separate preparations, or combined into a single composition for administration. Alternatively, the UPase inhibitor and the UR active agent can be administered sequentially to the subject via different preparations. Whether the UPase inhibitor and the UR active agent are administered sequentially or simultaneously, or in any effective variation thereof, for the purposes of the present invention, the agents are considered to be administered together or in combination. The routes of administration of the two agents may vary. Representative routes of administration are described below.

[0118] The subject treated according to the method of the present invention can be a subject suffering from or suspected of suffering from a liver disorder (e.g., NAFLD, NASH, or DILI). The subject at risk of liver disease or diagnosed with a severe liver disorder can be preventively started with the treatment according to the disclosed method. The treatment can be carried out at a more appropriate interval determined by a person skilled in the art. For example, administration can be carried out 1, 2, 3, 4 times or more per day. Ideally, the treatment is expected to be qd, for a long time. The treatment can also be started before, at the same time, or almost at the same time as the administration of the drug related to the severe liver disorder.

[0119] In the context of the present invention, the dose administered to an animal, especially a human, should be sufficient to affect a preventive or therapeutic response in the animal within a reasonable time frame. Those skilled in the art will recognize that the dose must depend on a variety of factors, including the specification of the specific compound used, the dosing regimen adopted, the animal's condition and the animal's weight, as well as the severity of the disease and the stage of the disease. The size of the dose may also be determined by the presence, nature and extent of any adverse side effects that may accompany the administration of a specific compound.

[0120] UPase inhibitors

[0121] As described above, aspects of the invention include administering a UPase inhibitor to the subject.UPase (UPh; EC 2.4.2.3) is a member of the pyrimidine nucleoside phosphorylase family, which catalyzes the phosphorylation cleavage of the CN glycosidic bond of UR to form ribose 1-phosphate and uracil. 45 The UPase inhibitor is an agent for regulating the plasma uridine level of the subject, for example, an agent for increasing the plasma uridine (UR) level of the subject. The increase in any plasma UR level may vary, and in some cases, the increase is 2-fold or higher, for example 5-fold or higher, 10-fold or higher, 15-fold or higher, 20-fold or higher, 25-fold or higher or 50-fold or higher.

[0122] In some cases, the UPase inhibitor is an anhydronucleoside. Anhydronucleoside is an analog of a natural nucleoside, which is commonly used as an intermediate for synthesizing nucleoside derivatives. They are characterized in that, except for having an N-glycosidic bond, they also have a covalent bond that is directly or through a bridging atom between a 2', 3' or 5' carbon atom of the sugar and a carbon, oxygen or nitrogen atom (except the nitrogen of the glycosidic bond) of the base. Anhydropyrimidine is characterized in that a pyrimidine base is covalently linked to a bridging atom between a 2', 3' or 5' carbon atom of the sugar and a carbon, oxygen or nitrogen atom (except the nitrogen of the glycosidic bond) of the pyrimidine base.

[0123] In some cases, the UPase inhibitor is 2,2'-anhydropyrimidine or a derivative thereof. In some embodiments, the 2,2'-anhydropyrimidine or a derivative thereof is a compound represented by formula (I):

[0124]

[0125] or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof;

[0126] in:

[0127] Each R 1 , R 2 , R 3 and R 4 are independently selected from the group consisting of hydrogen, substituted or unsubstituted heteroatoms, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, hydroxyl, halogen, azido, amino, substituted amino, carbohydrates, nucleic acids, amino acids, peptides, dyes, fluorophores and polypeptides.

[0128] In certain embodiments, the compound has a structural formula as shown in formula (I), R 1 , R 2 , R 3 and R 4 are independently hydrogen, hydroxyl, heteroatom, C1-C 18 Alkyl, C1-C 18 Substituted alkyl, C1-C 18 Alkenyl, C1-C 18 Acyl, amino, substituted amino, wherein the alkyl, alkenyl or acyl is a straight or branched chain group and is optionally substituted by hydroxy, ester and its derivatives, carboxyl and its derivatives, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, heteroatoms and may contain in-chain or bridging heteroatoms such as nitrogen, oxygen and sulfur.

[0129] Purpose 1Examples of components include, but are not limited to, hydrogen; hydroxyl; mercapto; halogens such as fluorine, chlorine, bromine or iodine, and pseudohalogens such as lower alkylsulfonyl groups having 1 to 5 carbon atoms (e.g., methyl-, ethyl-, propyl-, isopropyl-, butyl-, isobutyl-, tert-butyl-) and pentylsulfonyl or arylsulfonyl groups (e.g., benzene, p-toluene, p-nitrobenzenesulfonyl groups); lower alkyl groups having 1 to 20 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, etc., including substituted lower alkyl groups such as aminomethyl, hydroxymethyl, methoxy, ethoxy, propoxy, benzyloxy, imidoesters, alkylthio, (substituted substituted alkyl)thio, arylthio, (substituted aryl)thio, etc.; lower alkenyl containing 1 to 20 carbon atoms, such as vinyl and substituted vinyl, ethynyl and substituted ethynyl, wherein substituted vinyl or substituted ethynyl means that the β position of vinyl or ethynyl is substituted by halogen (such as bromine, chlorine, fluorine or iodine), or by alkyl having 1 to 5 carbon atoms (such as methyl, ethyl, propyl, butyl, pentyl, etc.), or aralkyl, such as benzyl, p-chlorobenzyl, p-nitrobenzyl, etc., or aryl, such as phenyl, p-nitrophenyl, p-tolyl, p-anisyl, naphthyl, etc.; lower alkanoyl (acyl) containing 1 to 20 carbon atoms substituted with 1 to 20 carbon atoms, such as phenyl, p-tolyl, p-chlorophenyl, p-aminophenyl, p-nitrophenyl, p-anisyl, etc.; substituted with 1 to 20 carbon atoms, such as benzoyl and naphthoyl, wherein the aromatic group may be further substituted with the following groups: alkyl, alkoxy, halogen or nitro moiety. or another aromatic acyl group, such as benzyloxybenzoyl, etc.; a lower aralkyl group containing 1 to 20 carbon atoms, such as benzyl, diphenylmethyl, p-chlorobenzyl, m-chlorobenzyl, p-nitrobenzyl, benzyloxybenzyl, pentafluorobenzyl, etc.; an amino group or an alkylamino group containing 1 to 20 carbon atoms, such as monoalkyl- or monoaralkylamino, such as methylamino, ethylamino, propylamino or benzylamino, etc.; a dialkylamino group, such as dimethylamino, diethylamino, dibenzylamino, pyrrolidino, piperidino or morpholino, etc.

[0130] Therefore, in certain embodiments, R 1 is hydrogen, hydroxyl, thiol, amino, substituted amino, hydroxymethyl, monomethoxy, halogen, pseudohalogen, or a lower hydrocarbon containing 1 to 20 atoms (wherein the hydrocarbon may be substituted or unsubstituted). In a specific embodiment, R1 is a lower hydrocarbon selected from alkyl, substituted alkyl, alkenyl, alkanoyl, aryl, aroyl, aralkyl or alkylamino. In a specific embodiment, R 1 is a lower hydrocarbon substituted by an alkoxy group, a substituted alkoxy group, an imido group, an arylthio group or a (substituted aryl)thio group. 1 is a lower alkyl group selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl and pentyl. 1 is a lower alkenyl selected from vinyl, substituted vinyl, ethynyl or substituted ethynyl. In other embodiments, R 1 is a lower alkanoyl selected from formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, tert-butyryl, valeryl, pivaloyl, caproyl, decanoyl, lauryl, myristyl, palmityl, stearoyl, eicosyl, stilligyl, palmitoyl, oleyl, linoleyl and arachidonic acid. In other embodiments, R 1 is a lower aryl group selected from phenyl, p-tolyl, p-chlorophenyl, p-aminophenyl, p-nitrophenyl, and p-anisyl. In other embodiments, R 1 is a lower aromatic acyl group selected from benzoyl and naphthoyl. 1 is a lower aralkyl group selected from benzyl, diphenylmethyl, p-chlorobenzyl, m-chlorobenzyl, p-nitrobenzyl, benzyloxybenzyl or pentafluorobenzyl. 1 It is a lower alkylamino group selected from the group consisting of a monoalkylamino group, a monoaralkylamino group, a dialkylamino group, a diaralkylamino group and a benzylamino group.

[0131] The target compound includes but is not limited to the compound represented by formula (I), wherein R 1 is selected from hydrogen, fluorine, trifluoromethyl, methyl, ethyl, propyl, butyl, isopropyl, isobutyl, acetyl, propionyl, butyryl, 2-bromovinyl, phenyl, benzyl, benzoyl, benzyloxybenzyl, benzylamino, alkoxyalkyl, benzyloxyalkyl, imidoalkyl ester, arylthio and (substituted aryl)thio. Therefore, in certain embodiments, the structural formula of the compound is as shown in formula (I), and R 1 It is H, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, benzyl, benzoyl, benzyloxybenzyl, benzyl-NH-, CH3CH2OCH2, benzyl-O-CH2, CH3OCH2, CH3C(NH)-O-CH2 or CH3-phenyl-O-CH2.

[0132] Purpose 2Examples of components include, but are not limited to: hydrogen; hydroxyl; mercapto; halogens such as fluorine, chlorine, bromine or iodine, and pseudohalogens such as lower alkylsulfonyl groups having 1 to 5 carbon atoms (e.g., methyl-, ethyl-, propyl-, isopropyl-, butyl-, isobutyl-, tert-butyl-) and pentylsulfonyl or arylsulfonyl groups (e.g., benzene, p-toluene, p-nitrobenzenesulfonyl groups); lower alkyl groups having 1 to 20 carbon atoms such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, etc., including substituted lower alkyl groups such as aminomethyl, hydroxymethyl, methoxy, ethoxy, propoxy, etc.; lower alkenyl groups having 1 to 20 carbon atoms such as vinyl and Substituted vinyl, ethynyl and substituted ethynyl, wherein substituted vinyl or substituted ethynyl means that the β position of vinyl or ethynyl is substituted by halogen (e.g., bromine, chlorine, fluorine or iodine), or by alkyl having 1 to 5 carbon atoms (e.g., methyl, ethyl, propyl, butyl, pentyl, etc.), or aralkyl, such as benzyl, p-chlorobenzyl, p-nitrobenzyl, etc., or aryl, such as phenyl, p-nitrophenyl, p-tolyl, p-anisyl, naphthyl, etc.; lower alkanoyl (acyl group) having 1 to 20 carbon atoms in the main chain and its esters, such as formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, tert-butyryl, valeryl, pivaloyl, hexanoyl, Decanoyl, lauryl, myristyl, palmityl, stearoyl, eicosyl, stilligyl, palmitoyl, oleyl, linolenoyl, arachidonic acid, etc.; lower aromatic groups containing 1 to 20 carbon atoms, such as phenyl, p-tolyl, p-chlorophenyl, p-aminophenyl, p-nitrophenyl, p-anisyl, etc.; lower aromatic groups containing 1 to 20 carbon atoms, such as benzoyl and naphthoyl, wherein the aromatic groups may be further substituted with the following groups: alkyl, alkoxy, halo or nitro moieties, such as p-tolnoyl, p-anisyl, p-chlorobenzoyl, p-nitrobenzoyl or 2,4-dinitrobenzoyl, pentafluorobenzoyl, etc., or another a 4-(4-chlorobenzyl)-oxy group (i.e., O-benzyl)-1,4-(4-(4-chlorobenzyl)-1,4-(4-(4-chlorobenzyl)-1),4-(4-(4-benzyloxy)-oxy group (i.e., O-benzyl)-1,4-(4-(4-chlorobenzyl)-1),4-(4-(4-chlorobenzyl ...chlorobenzyl)-1),4-(4-chlorobenzEsters of aryloxy groups, for example, lower aroyloxy groups (i.e., O-aroyl groups) containing 1 to 20 carbon atoms, such as benzoyloxy groups (i.e., O-benzoyl groups), diphenylacetyloxy groups (i.e., O-diphenylacetyl groups), p-chlorobenzoyloxy groups (i.e., O-(p-chlorobenzoyl) groups), m-chlorobenzoyloxy groups (i.e., O-(m-chlorobenzoyl) groups), p-nitrobenzoyloxy groups (i.e., O-(p-nitrobenzoyl) groups), (4-benzyloxybenzoyl)-oxy groups (i.e., O-benzyloxybenzoyl) groups, or pentafluorobenzoyloxy groups (i.e., O-pentafluorobenzoyl groups); amino groups or alkylamino groups containing 1 to 20 carbon atoms, such as monoalkyl- or monoaralkylamino groups, such as methylamino, ethylamino, propylamino, or benzylamino groups, etc.; dialkylamino groups, such as dimethylamino, diethylamino, dibenzylamino, pyrrolidino, piperidino, or morpholino groups, etc.;

[0133] Therefore, in certain embodiments, R 2 is hydrogen, hydroxyl, mercapto, amino, hydroxymethyl, monomethoxy, halogen, pseudohalogen, or a lower hydrocarbon containing 1 to 20 atoms (wherein the hydrocarbon may be substituted or unsubstituted) and esters thereof. 2 is a lower hydrocarbon selected from alkyl, alkenyl, alkanoyl, aryl, aroyl, aryloxy, aroyloxy, aralkyl or alkylamino. 2 is a lower alkyl group selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl and pentyl. 2 is a lower alkenyl selected from vinyl, substituted vinyl, ethynyl or substituted ethynyl. In other embodiments, R 2 is a lower alkanoyl selected from formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, tert-butyryl, valeryl, pivaloyl, caproyl, decanoyl, lauryl, myristyl, palmityl, stearoyl, eicosyl, stilligyl, palmitoyl, oleyl, linoleyl and arachidonic acid. In other embodiments, R 2 is a lower aryl group selected from phenyl, p-tolyl, p-chlorophenyl, p-aminophenyl, p-nitrophenyl, and p-anisyl. In other embodiments, R 2 is a lower aromatic acyl group selected from benzoyl and naphthoyl. 2 is a lower aralkyl group selected from benzyl, diphenylmethyl, p-chlorobenzyl, m-chlorobenzyl, p-nitrobenzyl, benzyloxybenzyl or pentafluorobenzyl. 2 is a lower aryloxy group selected from phenoxy, benzyloxy, benzhydryloxy, p-chlorobenzyloxy, m-chlorobenzyloxy, p-nitrobenzyloxy, (4-benzyloxybenzyl)-oxy or pentafluorobenzyloxy. 2is a lower aroyloxy group selected from benzoyloxy, diphenylacetyloxy, p-chlorobenzoyloxy, m-chlorobenzoyloxy, p-nitrobenzoyloxy, (4-benzyloxybenzoyl)-oxy or pentafluorobenzoyloxy. In certain other embodiments, R 2 is a lower alkylamino group selected from monoalkylamino, monoaralkylamino, dialkylamino and diaralkylamino. 2 In addition to hydrogen or hydroxy, O-acyl, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, O-alkyl, O-alkylene, O-alkynyl, O-aralkyl, O-aryl, O-aryloxy, O-carbohydrate, O-cycloalkenyl, O-cycloalkyl, O-heterocycloalkyl, O-heteroaryl can also be used. In addition, O can be replaced by S.

[0134] The target compound includes but is not limited to the compound represented by formula (I), wherein R 2 is selected from hydrogen, fluorine, trifluoromethyl, methyl, ethyl, propyl, butyl, isopropyl, isobutyl, acetyl, propionyl, butyryl, 2-bromovinyl, phenyl, phenoxy, benzyl, benzoyl, benzoyloxy and benzyloxybenzyl. Therefore, in certain embodiments, the structural formula of the compound is as shown in formula (I), and R 2 It is H, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, phenoxy, benzyl, benzoyl, benzoyloxy or benzyloxybenzyl.

[0135] In a specific embodiment, the compound has a structural formula as shown in formula (I), and R 2 is hydrogen, hydroxyl or a substituent connected to O. This includes compounds represented by formula (I) wherein R 2 It is H, OH or C5H5C(O)O.

[0136] Purpose 3Examples of include, but are not limited to, hydrogen; hydroxyl; azido; mercapto; halogen; pseudohalogen; lower alkyl groups containing 1 to 20 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, etc., including substituted lower alkyl groups, such as aminomethyl, hydroxymethyl, methoxy, ethoxy, propoxy, etc.; lower alkanoyl (acyl) groups having a main chain of 1 to 20 carbon atoms, including esters thereof, such as formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, tert-butyryl, valeryl, pivaloyl, hexanoyl, decanoyl, lauryl, myristyl, palmityl, stearyl, eicosyl, stilligyl, palmitoyl, oleyl, linoleyl, arachidonic acid, etc.; lower aryl groups, such as phenyl, p-nitrophenyl, p-tolyl, p-anisyl, naphthyl, etc.; A lower aromatic acyl group (acyl group of an aromatic acid) having 1 to 20 carbon atoms, such as benzoyl and naphthoyl, wherein the aromatic group may be further substituted with an alkyl, alkoxy, halo, or nitro moiety, such as p-tolnoyl, p-anisyl, p-chlorobenzoyl, p-nitrobenzoyl or 2,4-dinitrobenzoyl, pentafluorobenzoyl, etc.; a lower aromatic oxy group having 1 to 20 carbon atoms, such as phenoxy, benzyloxy, diphenylmethoxy, p-chlorobenzyloxy, m-chlorobenzyloxy, p-nitrobenzyloxy, (4-benzyloxybenzyl)-oxy or pentafluorobenzyloxy, etc.; and an ester of an aromatic oxy group, such as a lower aromatic acyloxy group (O-aroyl) having 1 to 20 carbon atoms, such as benzoyloxy, diphenylacetyloxy, p-chlorobenzoyloxy, m-chlorobenzoyloxy, p-nitrobenzoyloxy, (4-benzyloxybenzoyl)-oxy or pentafluorobenzoyloxy, etc. 3 It may also be an adamantanoyl group or a substituted adamantanoyl group.

[0137] Therefore, in certain embodiments, R 3 is hydrogen, hydroxyl, azido, thiol, hydroxymethyl, halogen or pseudohalogen. 3 is a lower hydrocarbon selected from alkyl, alkanoyl, aryl, aroyl, aryloxy, aroyloxy or aralkyl. 3 is a lower alkyl group selected from methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl and pentyl. 3 is a lower alkanoyl selected from formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, tert-butyryl, valeryl, pivaloyl, caproyl, decanoyl, lauryl, myristyl, palmityl, stearoyl, eicosyl, stilligyl, palmitoyl, oleyl, linoleyl and arachidonic acid. In other embodiments, R 3is a lower aryl group selected from phenyl, p-tolyl, p-chlorophenyl, p-aminophenyl, p-nitrophenyl, p-anisyl, and the like. In other embodiments, R 3 is a lower aromatic acyl group selected from benzoyl and naphthoyl. In other certain embodiments, R 3 is a lower aralkyl group selected from benzyl, diphenylmethyl, p-chlorobenzyl, m-chlorobenzyl, p-nitrobenzyl, benzyloxybenzyl or pentafluorobenzyl. 3 is a lower aryloxy group selected from phenoxy, benzyloxy, benzhydryloxy, p-chlorobenzyloxy, m-chlorobenzyloxy, p-nitrobenzyloxy, (4-benzyloxybenzyl)-oxy or pentafluorobenzyloxy. 3 is a lower aroyloxy group selected from benzoyloxy, diphenylacetyloxy, p-chlorobenzoyloxy, m-chlorobenzoyloxy, p-nitrobenzoyloxy, (4-benzyloxybenzoyl)-oxy or pentafluorobenzoyloxy. 3 In addition to hydrogen or hydroxy, O-acyl, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, O-alkyl, O-alkylene, O-alkynyl, O-aralkyl, O-aryl, O-aryloxy, O-carbohydrate, O-cycloalkenyl, O-cycloalkyl, O-heterocycloalkyl, O-heteroaryl can also be used. In addition, O can be replaced by S.

[0138] The target compound is a compound represented by formula (I), wherein R 3 is hydrogen, hydroxyl, halogen, azido or an O-linked substituent. This includes compounds of formula (I) wherein R 3 is selected from hydrogen, hydroxy, n-butoxy, isobutoxy, tert-butoxy, phenoxy, benzyloxy, benzoyloxy and pentafluorobenzoyloxy. Therefore, in certain embodiments, the structural formula of the compound is as shown in formula (I), and R 3 Selected from H, OH, CH3CH2CH2CH2O, (CH3)2CH2CH2O, (CH3)3CO, C6H5O, benzoyloxy and pentafluorobenzoyloxy.

[0139] In a specific embodiment, the structural formula of the compound is as shown in formula (I), wherein R 3 is H, OH, F, Cl, Br, I, N3 or C5H5C(O)O. Of particular interest are compounds of formula (I) wherein R3 is OH or O-acyl (e.g., an ester, such as C5H5C(O)O).

[0140] R 4 Examples of R include, but are not limited to: hydrogen; hydroxy; mercapto; halogen, such as fluorine, chlorine, bromine or iodine; amino or lower alkylamino. 4Also exemplified are lower alkyl groups containing acyl groups, which may be lower alkanoyl groups having 1 to 7 carbon atoms, such as formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, tert-butyryl, and the like, and esters thereof. 4 It can also be an aromatic acyl group (and its ester, such as an aromatic acyl group connected to O, i.e., O-aroyl or aromatic acyloxy), such as benzoyl and naphthoyl, wherein the aromatic group can be further substituted with the following groups: alkyl, alkoxy, halo or nitro moieties, such as p-tolnoyl, p-anisyl, p-chlorobenzoyl, p-nitrobenzoyl or 2,4-dinitrobenzoyl, etc. Therefore, in certain embodiments, R 4 In addition to hydrogen or hydroxy, O-acyl, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, O-alkyl, O-alkylene, O-alkynyl, O-aralkyl, O-aryl, O-aryloxy, O-carbohydrate, O-cycloalkenyl, O-cycloalkyl, O-heterocycloalkyl, O-heteroaryl can also be used. In addition, O can be replaced by S.

[0141] Therefore, in certain embodiments, R 4 is hydrogen; hydroxyl; thiol; halogen, aminoaminomethyl or aminodimethyl. In other embodiments, R 4 is a lower alkyl, acyl, aroyl or aroyloxy group. This includes specific embodiments in which the compound represented by formula (I) is a compound in which R4 is hydrogen, fluorine, hydroxyl, amino, aminomethyl, aminodimethyl, tert-butoxy, phenoxy or benzoyloxy (e.g., a compound represented by formula (I) in which R 4 It is H, F, OH, NH2, NHCH3, N(CH3) 2、 (CH3)3CO, C6H5O or C6H5C(O)O).

[0142] The specific target compound is a compound represented by formula (I), wherein R 4 is hydrogen, hydroxyl or a substituent connected to O. In a specific embodiment, the structural formula of the compound is as shown in formula (I), wherein R 4 is H, OH or C6H5C(O)O. Of particular interest are compounds of formula (I) wherein R 4 is OH or O-acyl (eg, an ester, such as C6H5C(O)O).

[0143] Of interest are compounds of formula (I), wherein: R 1 is H, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, benzyl, benzoyl or benzyloxybenzyl, R 2is H, OH, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, phenoxy, benzyl, benzoyl, benzoyloxy or benzyloxybenzyl, and wherein R 3 and R 4 These compounds include the following: 2,2'-anhydrouridine; 2,2'-anhydro-5-fluorouridine; 2,2'-anhydro-5-trifluoromethyluridine; 2,2'-anhydro-5-methyluridine; 2,2'-anhydro-5-ethyluridine; 2,2'-anhydro-5-propyluridine; 2,2'-anhydro-5-isopropyluridine; 2,2'-anhydro-5-isobutyluridine; 2,2'-anhydro-5-methylacyluridine; 2,2'-anhydro-5-propylacyluridine; 2,2'-anhydro-5-(2-bromovinyl)-uridine; 2,2'-anhydro-5-phenyluridine; 2,2'-anhydro-5-benzyluridine; 2,2'-anhydro-5-benzoyluridine; and 2,2'-anhydro-5-(benzyloxybenzyl)-uridine. Of particular interest is 2,2'-anhydro-5-methyluridine or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof.

[0144] Other target compounds are compounds represented by formula (I), wherein: R 1 is H, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, benzyl, benzoyl or benzyloxybenzyl, R 2 is H, OH, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, phenoxy, benzyl, benzyloxy, benzoyl, benzoyloxy or benzyloxybenzyl, and wherein R 3 is hydroxyl, R 4These compounds include the following: 3'-O-benzoyl-2,2'-anhydrouridine; 3'-O-benzoyl-2,2'-anhydro-5-fluorouridine; 3'-O-benzoyl-2,2'-anhydro-5-trifluoromethyluridine; 3'-O-benzoyl-2,2'-anhydro-5-methyluridine; 3'-O-benzoyl-2,2'-anhydro-5-ethyluridine; 3'-O-benzoyl-2,2'-anhydro-5-propyluridine; 3'-O-benzoyl-2,2'-anhydro-5-isopropyluridine; 3'-O-benzoyl-2,2'-O-anhydro-5- -5-isobutyluridine; 3'-O-benzoyl-2,2'-anhydro-5-methylacyluridine; 3'-O-benzoyl-2,2'-anhydro-5-propylacyluridine; 3'-O-benzoyl-2,2'-anhydro-5-(2-bromovinyl)-uridine; 3'-O-benzoyl-2,2'-anhydro-5-phenyluridine; 3'-O-benzoyl-2,2'-anhydro-5-benzyluridine; 3'-O-benzoyl-2,2'-anhydro-5-benzoyluridine; and 3'-O-benzoyl-2,2'-anhydro-5-(benzyloxybenzyl)-uridine. Of particular interest is 3'-O-benzoyl-2,2'-anhydro-5-methyluridine or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof.

[0145] Also of interest are compounds of formula (I), wherein: R 1 is H, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, benzyl, benzoyl or benzyloxybenzyl, R 2 is H, OH, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, phenoxy, benzyl, benzyloxy, benzoyl, benzoyloxy or benzyloxybenzyl, and wherein R 3 is benzoyloxy, R 4These compounds include the following: 5'-O-benzoyl-2,2'-anhydrouridine; 5'-O-benzoyl-2,2'-anhydro-5-fluorouridine; 5'-O-benzoyl-2,2'-anhydro-5-trifluoromethyluridine; 5'-O-benzoyl-2,2'-anhydro-5-methyluridine; 5'-O-benzoyl-2,2'-anhydro-5-ethyluridine; 5'-O-benzoyl-2,2'-anhydro-5-propyluridine; 5'-O-benzoyl-2,2'-anhydro-5-isopropyluridine; 5'-O-benzoyl-2,2'-O-anhydro-5- -5-isobutyluridine; 5'-O-benzoyl-2,2'-anhydro-5-methylacyluridine; 5'-O-benzoyl-2,2'-anhydro-5-propylacyluridine; 5'-O-benzoyl-2,2'-anhydro-5-(2-bromovinyl)-uridine; 5'-O-benzoyl-2,2'-anhydro-5-phenyluridine; 5'-O-benzoyl-2,2'-anhydro-5-benzyluridine; 5'-O-benzoyl-2,2'-anhydro-5-benzoyluridine; and 5'-O-benzoyl-2,2'-anhydro-5-(benzyloxybenzyl)-uridine. Of particular interest is 5'-O-benzoyl-2,2'-anhydro-5-methyluridine or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof.

[0146] The 2,2'-anhydropyrimidine compound of the present invention can be included in a composition containing a single stereoisomer, a mixture of stereoisomers, and a variety of derivatives thereof that can be produced as an equilibrium mixture of tautomers. For example, the 2,2'-anhydropyrimidine according to formula (I) includes four stereocenters on the furanocyclic ring, and the ring includes α and β end group isomers, as well as L or D mirror configurations. Examples of stereoisomers of the 2,2'-anhydropyrimidine compound of the present invention are β-D-isomers, β-L-isomers, α-D-isomers, α-L-isomers, and tautomers and mixtures including α,-β-D-isomers, α,β-L-isomers, α-DL-isomers, and β-DL-isomers. Thus, in one embodiment, a composition consisting essentially of a stereoisomer of 2,2'-anhydropyrimidine that is a β-D-isomer, a β-L-isomer, an α-D-isomer, or an α-L-isomer is provided.

[0147] Specific target stereoisomers include: 2,2'-anhydro-1-(β-D-arabinofuranosyl) uracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-fluorouracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-n-propyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyl ...trifluoromethyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl) uracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isobutyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-formyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-propionyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-phenyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzyluracil; 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzoyluracil; and 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(3-benzyloxybenzyl)uracil.Further stereoisomers of interest include: 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl) uracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-fluorouracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-n-propyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-n-propyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil; 3'-O-benzoyl-2,2 '-anhydro-1-(β-D-arabinofuranosyl)-5-phenyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzyluracil; 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzoyluracil; and 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(3-benzyloxybenzyl)uracil.Other stereoisomers of interest include: 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl) uracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-fluorouracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-n-propyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-n-propyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-formyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-propylacyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil; 5'-O-benzoyl-2,2 '-anhydro-1-(β-D-arabinofuranosyl)-5-phenyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzoyluracil; and 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(3-phenoxybenzyl)uracil.

[0148] Examples of other analogs or derivatives of 2,2'-anhydropyrimidine and its stereoisomers of the present invention include: 3'-O-acetyl-2,2'-anhydro-5-propyluridine (3'-O-acetyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-propyluracil); 3'-O-acetyl-2,2'-anhydro-5-isopropyluridine (3'-O-acetyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyluracil); and 2,2'-anhydrocytidine and its analogs and derivatives, wherein the stereoisomer 2,2'-anhydro-1-(β-D-arabinofuranosyl)cytosine is an example.

[0149] As described above, the specific target stereoisomers and various 2,2'-anhydropyrimidines are stereoisomers that show improved activity on a molar basis. For this reason, such compounds can be easily selected by comparing the matrix of the specific target compound, such as the compounds exemplified in Table 4 (wherein the structural formula of the compound is shown in Formula (I)).

[0150]

[0151]

[0152] As mentioned above, the compounds in Table 4 are for illustration only and are not intended to be limiting. 4 In addition to hydroxyl, O-acyl, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, O-alkyl, O-alkylene, O-alkynyl, O-aralkyl, O-aryl, O-aryloxy, O-carbohydrate, O-cycloalkenyl, O-cycloalkyl, O-heterocycloalkyl, O-heteroaryl can also be used. In addition, S can replace O, and other combinations of the structural elements (for example, as described herein) and other stereochemical orientations are also possible.

[0153] In some embodiments, the acyl derivatives of 2,2'-anhydropyrimidine of formula (I) are of interest. 1 , R 2 , R 3 and R 4 A compound as defined above, wherein R 2 , R 3 and R 4 At least one of them is an acyl derivative. "Acyl derivative" means a derivative of 2,2'-anhydropyrimidine represented by formula (I), wherein R 2 , R 3 and R 4 At least one of the substituents is a substantially non-toxic organic acyl substituent derived from a carboxylic acid, and the carboxylic acid is linked to the hydroxyl group on the ribose or pyrimidine ring of formula (I) through an ester bond.

[0154] The acyl derivatives of the 2,2'-anhydropyrimidine compounds represented by formula (I) include those wherein R 1 As defined above, and each R 2 , R 3 and R 4 are independently hydrogen, hydroxyl or acyl, provided that R 2 , R 3 and R 4 In another embodiment, the acyl derivative of 2,2'-anhydropyrimidine is a compound represented by formula (I), wherein R1 and R 2 As defined above, the premise is that R 2 Not hydrogen, each R 3 and R 4 In one embodiment, the acyl derivative of 2,2'-anhydropyrimidine is a compound represented by formula (I), wherein R 1 As defined above, R 2 is hydrogen, and each R 3 and R 4 are independently hydroxyl or acyl. Of particular interest are acyl derivatives of 2,2'-anhydropyrimidine compounds of formula (I), wherein R 1 is methyl, R 2 is hydrogen, and each R 3 and R 4 Also of interest are acyl derivatives of 2,2'-anhydropyrimidine compounds of formula (I), wherein R 1 is methyl, R 2 is hydrogen, and each R 3 and R 4 are independently acyl.

[0155] Typically, the ester bond of the acyl derivative shown in formula (I) can be cleaved under physiological conditions, in vitro (e.g., cell-based systems) and / or in vivo (e.g., by body metabolism). Therefore, in certain embodiments, the acyl group is a group of a metabolite. Such acyl substituents include, but are not limited to, substituents derived from the following acids: acetic acid, fatty acids, amino acids, lipoic acid, glycolic acid, lactic acid, enolpyruvic acid, pyruvic acid, orotic acid, acetoacetic acid, β-hydroxybutyric acid, creatine, succinic acid, fumaric acid, adipic acid, benzoic acid, and p-aminobenzoic acid. Specific acyl substituents of interest are compounds that are normally present in the body as dietary components or intermediate metabolites, and when cleaved from the target 2,2'-anhydropyrimidine compound in vivo, the compound is substantially non-toxic.

[0156] Of particular interest are compositions comprising 3'-O-acyl-2,2'-anhydropyrimidine or derivatives thereof. For example, the acyl derivative of interest is a compound comprising a 2,2'-anhydropyrimidine compound of formula (I), wherein each R 1 , R 2 and R 3 are independently selected from hydrogen, hydroxy, mercapto, amino, hydroxymethyl, methoxy, halogen, pseudohalogen and substituted or unsubstituted lower hydrocarbons containing 1 to 20 carbon atoms, such as lower hydrocarbons selected from alkyl, alkenyl, alkanoyl, aryl, aroyl, aralkyl and alkylamino, and esters thereof, and wherein R 4 It is an O-acyl group.

[0157] In certain embodiments, the acyl derivative comprises a 2,2'-anhydropyrimidine compound represented by formula (I), wherein R 4 is an O-acyl group, and wherein the O-acyl group contains 1 to 10 carbon atoms, for example, an O-acyl group selected from aroyloxy, aralkanoyloxy, heteroaroyloxy and cycloalkanoyloxy.

[0158] Therefore, the acyl derivatives of the 2,2'-anhydropyrimidine compound shown in formula (I) include 3'-O-acyl-2,2'-anhydropyrimidine, 5'-O-acyl-2,2'-anhydropyrimidine, 3',5'-O-acyl-2,2'-anhydropyrimidine and derivatives thereof. For example, 3'-O-acyl-2,2'-anhydropyrimidine or derivatives thereof include 3'-O-aroyl-2,2'-anhydropyrimidine, such as 3'-O-aroyl-2,2'-anhydrouridine or derivatives thereof. An example of the derivative of particular interest is 3'-O-benzoyl-2,2'-anhydrouridine or derivatives thereof, such as 3'-O-benzoyl-2,2'-anhydro-5-methyluridine. Of interest are compounds in which 3'-O-benzoyl-2,2'-anhydro-5-methyluridine is the stereoisomer 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil.

[0159] In some embodiments, the acyl derivatives of the 2,2'-anhydropyrimidine compound of formula (I) include compounds satisfying the following conditions, wherein: R 1 is H, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, benzyl, benzoyl or benzyloxybenzyl, R 2 is H, OH, F, CF3, CH3, CH3CH2, CH3CH2CH2, (CH3)2CH, (CH3)2CH2CH2, CH3(O)CCH2, CH3(O)CCH2CH2, Br-CH=CH, phenyl, phenoxy, benzyl, benzyloxy, benzoyl, benzyloxybenzyl or acyl, and wherein each R 3 and R 4are independently hydroxyl or acyl. These compounds include the following compounds: 3'-O-benzoyl-2,2'-anhydrouridine; 3'-O-benzoyl-2,2'-anhydro-5-fluorouridine; 3'-O-benzoyl-2,2'-anhydro-5-trifluoromethyluridine; 3'-O-benzoyl-2,2'-anhydro-5-methyluridine; 3'-O-benzoyl-2,2'-anhydro-5-ethyluridine; 3'-O-benzoyl-2,2'-anhydro-5-propyluridine; 3'-O-benzoyl-2,2'-anhydro-5-isopropyluridine; 3'-O-benzoyl-2,2'-O-anhydro-5-isobutyluridine; 3'-O-benzoyl-2,2'-anhydro-5-formyluridine; 3' 3'-O-benzoyl-2,2'-anhydro-5-propionyl uridine; 3'-O-benzoyl-2,2'-anhydro-5-(2-bromovinyl)-uridine; 3'-O-benzoyl-2,2'-anhydro-5-phenyl uridine; 3'-O-benzoyl-2,2'-anhydro-5-benzyl uridine; 3'-O-benzoyl-2,2'-anhydro-5-benzoyl uridine; and 3'-O-benzoyl-2,2'-anhydro-5-(benzyloxybenzyl)-uridine; 5'-O-benzoyl-2,2'-anhydro uridine; 5'-O-benzoyl-2,2'-anhydro-5-fluoro uridine; 5'-O-benzoyl-2,2'-anhydro-5-trifluoromethyl ... 5'-O-benzoyl-2,2'-anhydro-5-methyluridine; 5'-O-benzoyl-2,2'-anhydro-5-ethyluridine; 5'-O-benzoyl-2,2'-anhydro-5-propyluridine; 5'-O-benzoyl-2,2'-anhydro-5-isopropyluridine; 5'-O-benzoyl-2,2'-anhydro-5-isobutyluridine; 5'-O-benzoyl-2,2'-anhydro-5-formyluridine; 5'-O-benzoyl-2,2'-anhydro-5-propionyluridine; 5'-O-benzoyl-2,2'-anhydro-5-(2-bromovinyl)-uridine; 5'-O-benzoyl-2,2'-anhydro-5-phenyluridine; 5'-O-benzoyl 5'-O-benzoyl-2,2'-anhydro-5-benzyl uridine; 5'-O-benzoyl-2,2'-anhydro-5-benzoyl uridine; and 5'-O-benzoyl-2,2'-anhydro-5-(benzyloxybenzyl)-uridine; 3',5'-O-benzoyl-2,2'-anhydro uridine; 3',5'-O-benzoyl-2,2'-anhydro-5-fluoro uridine; 3',5'-O-benzoyl-2,2'-anhydro-5-trifluoromethyl uridine; 3',5'-O-benzoyl-2,2'-anhydro-5-methyl uridine; 3',5'-O-benzoyl-2,2'-anhydro-5-ethyl uridine; 3',5'-O-benzoyl-2,2'-anhydro-5-propyl uridine;3',5'-O-benzoyl-2,2'-anhydro-5-isopropyluridine; 3',5'-O-benzoyl-2,2'-O-anhydro-5-isobutyluridine; 3',5'-O-benzoyl-2,2'-anhydro-5-formyluridine; 3',5'-O-benzoyl-2,2'-anhydro-5-propionyluridine; 3',5'-O-benzoyl-2,2'-anhydro-5-(2-bromovinyl)-uridine; 3',5'-O-benzoyl-2,2'-anhydro-5-phenyluridine; 3',5'-O-benzoyl-2,2'-anhydro-5-benzyluridine; 3',5'-O-benzoyl-2,2'-anhydro-5-benzyluridine; and 3',5'-O-benzoyl-2,2'-anhydro-5-(benzyloxybenzyl)-uridine; or pharmaceutically acceptable salts, solvates, hydrates, prodrug forms thereof, and stereoisomers thereof. ;

[0160] Of particular interest are 3'-O-benzyl-2,2'-anhydro-5-methyluridine, 5'-O-benzyl-2,2'-anhydro-5-methyluridine and 3',5'-O-benzyl-2,2'-anhydro-5-methyluridine, or pharmaceutically acceptable salts, solvates, hydrates, prodrug forms thereof, and stereoisomers thereof. Of particular interest are the β-D-arabinofuranosyl isomers of these compounds or pharmaceutically acceptable salts, solvates, hydrates, and prodrug forms thereof.

[0161] In another embodiment, the compound according to the specific object formula (I) is wherein R 1 and R 4 As defined above, and R 2 and / or R 3 The compound is a cycloalkyl group. "Cycloalkyl" means a hydrocarbon ring structure having 3 to about 10 carbon atoms and having a single ring or multiple condensed rings, which may be substituted. The cycloalkyl group of interest is selected from aryl, aralkyl, aryloxy, aroyl, aroyloxy, heteroaryl, heteroaryloxy, heteroaroyloxy, cycloalkyl, cycloalkyloxy and cycloalkanoyloxy. Therefore, the cycloalkyl group of particular interest is a ribose or pyrimidine ring of formula (I) connected to O. Where R 2 and / or R 3 Compounds that are cycloalkyl show improved activity on a molar basis.

[0162] Thus, certain compounds of the present invention comprise 5'-O-(cycloalkyl)-2,2'-anhydropyrimidine or a derivative thereof. This embodiment includes 5'-O-(cycloalkyl)-2,2'-anhydro-5(R 5 )-uridine or its derivatives, wherein R 5 YesR 1 (For example, R 5 =R1 , where "5(R 5 )" refers to R 1 and the same).

[0163] The target compound is a 5'-O-aryl-2,2'-anhydropyrimidine or a derivative thereof, including various 2,2'-anhydrouridine derivatives. This includes compounds in which the 5'-O-aryl-2,2'-anhydropyrimidine is a 5'-O-aroyl-2,2'-anhydropyrimidine, such as: 5'-O-benzyl-2,2'-anhydropyrimidine; 5'-O-chlorobenzyl-2,2'-anhydropyrimidine; 5'-O-nitrobenzyl-2,2'-anhydropyrimidine; 5'-O-hydroxybenzyl-2,2'-anhydropyrimidine, etc.

[0164] In one embodiment, the compound that exhibits improved activity on a molar basis or that exhibits improved specificity in terms of not interfering with the efficacy of fluorouracil therapy is 5'-O-aryl-2,2'-anhydrouridine, 5'-O-aroyl-2,2'-anhydrouridine and derivatives thereof, such as 5'-O-aryl-2,2'-anhydro-5(R 4 )-uridine, 5'-O-aroyl-2,2'-anhydro-5(R 4 )-uridine and its derivatives. Examples include 5'-O-aryl-2,2'-anhydro-5-methyl-uridine; 5'-O-aryl-2,2'-anhydro-5-ethyl-uridine; 5'-O-aryl-2,2'-anhydro-5-propyl-uridine; 5'-O-aryl-2,2'-anhydro-5-benzyl-uridine; 5'-O-aryl-2,2'-anhydro-5-(2-bromovinyl)-uridine; and derivatives thereof. Examples also include 5'-O-aroyl-2,2'-anhydro-5-methyl-uridine; 5'-O-aroyl-2,2'-anhydro-5-ethyl-uridine; 5'-O-aroyl-2,2'-anhydro-5-propyl-uridine; 5'-O-aroyl-2,2'-anhydro-5-benzyl-uridine; 5'-O-aroyl-2,2'-anhydro-5-(2-bromovinyl)-uridine; and derivatives thereof. Specific compounds of interest include 5'-O-benzoyl-2,2'-anhydro-5(R 4 )-uridine, such as 5'-O-benzoyl-2,2'-anhydro-5-methyl-uridine; 5'-O-benzoyl-2,2'-anhydro-5-ethyl-uridine; 5'-O-benzoyl-2,2'-anhydro-5-propyl-uridine; 5'-O-benzoyl-2,2'-anhydro-5-benzyl-uridine; and 5'-O-benzoyl-2,2'-anhydro-5-(2-bromovinyl)-uridine.

[0165] The stereoisomers of interest include 5'-O-(cycloalkyl)-2,2'-anhydropyrimidines that are β-D-isomers. Examples include, but are not limited to: 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)uracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-fluorouracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-trifluoromethyluracil; -5-methyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-n-propyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-isopropyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-ethyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-formyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-propylacyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(2-bromovinyl)uracil -anhydro-1-(β-D-arabinofuranosyl)-5-phenyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzyluracil; 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-benzoyluracil; and 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-(3-phenoxybenzyl)uracil.

[0166] As mentioned above, also of interest are analogs / derivatives of the above compounds.

[0167] The above-mentioned 2,2'-anhydropyrimidines and their derivatives are commercially available or can be prepared in a conventional manner by techniques known to those skilled in the art. For example, representative patents describing various 2,2'-anhydropyrimidines and derivatives (including intermediates and precursors thereof, analysis, and synthesis / preparation thereof) include the following U.S. Patents: 3,975,367; 4,145,531; 4,230,698; 4,247,544; 4,544,740; 4,604,382; 4,613,604; 4,681,933; 4,841,039; 4,916,122; 4,987,224; 5,008,384; 5,077,280; 5,084,445; 5,141,943; 5,190,926; 5,212 The contents of these patents are incorporated herein by reference.

[0168] Uridine phosphorylase (UPase) inhibitors also include, but are not limited to: benzyl cyclic uridine, benzyloxy acyl uridine, aminomethyl-benzyl acyl uridine, aminomethyl-benzyloxybenzyl cyclic uridine, hydroxymethyl-benzyl cyclic uridine, hydroxymethyl-benzyloxybenzyl cyclic uridine, etc.; derivatives of 5-benzyl barbiturate, such as 5-benzyloxybenzyl barbiturate; 5-benzyloxybenzyl-1-(1-hydroxy-2-ethoxy)methyl) barbiturate; 5-benzyloxybenzyl acetyl-1-(1-hydroxy-2-ethoxy)methyl) barbiturate; 5-benzyloxybenzyl-1-(1,3-dihydroxy 2-propoxy)methyl barbiturate; 5- benzyloxybenzyl-1-(1-hydroxy, 3-amino-2-propoxy)methyl)barbiturate; 5-benzyloxybenzyl-1-(2-(3-carboxypropionyloxy)ethoxy)methyl)barbiturate; 5-benzyl-1-(1-hydroxy-2-ethoxy)methyl)barbiturate; 5-methoxybenzylacetyl barbiturate; 5-benzyl-1-(1,3-dihydroxy-2-propoxy)methyl)barbiturate; 5-benzyl-1-(1-hydroxy, 3-amino-2-propoxy)methyl)barbiturate; and 5-benzyl-1-(2-(3-(carboxypropionyloxy)ethoxy)methyl)barbiturate, etc. Upase inhibitors that can be used in embodiments of the present invention include, but are not limited to, those described in the following U.S. Patents: 5,723,449; 5,141,943; 5,077,280; and 4,613,604; the disclosures of the compounds disclosed in these publications are incorporated herein by reference.

[0169] Uridine (UR) active agent

[0170] As described above, in some embodiments, the UPase inhibitor is administered to the subject in combination with a UR active agent (e.g., uridine (UR), a UR prodrug, or a UR mimetic). Uridine is a nucleoside formed when uracil is linked to a ribose ring (also known as furanose) via a β-N1-glycosidic bond. Uridine is provided in phosphorylated form, i.e., uridine-5'-monophosphate (also known as 5'-uridylic acid and UMP), uridine 5'-monophosphate tris salt, uridine 5'-monophosphate salt dihydrate, uridine 5'-monophosphate salt solution, uridine 5'-monophosphate salt hydrate, uridine 13C 9. 15 N25'-monophosphate sodium salt solution, uridine- 15 N2 5'-monophosphate sodium salt solution, uridine 5'-monophosphate trisodium hydrate, uridine-N25'-monophosphate sodium salt solution, uridine-5'-diphosphate (UDP), uridine 5'-diphosphate trishydroxymethylaminomethane salt, uridine 5'-diphosphate salt dihydrate, uridine 5'-diphosphate salt solution, uridine 5'-diphosphate salt hydrate, uridine13C 9. 15 N25'-diphosphate sodium salt solution, uridine-5'-triphosphate (UTP), UTPγS, MRS2498, uridine 5'-triphosphate trishydroxymethylaminomethane salt, uridine 5'-triphosphate salt dihydrate, uridine 5'-triphosphate salt solution, uridine 5'-triphosphate salt hydrate, uridine 13C 9. 15 15 N25'-5'-triphosphate sodium salt solution, 2-douridine tetraphosphate, thio-UTP tetrasodium salt, denufosol tetrasodium or UTP-γ-S trisodium salt, a prodrug known in the art as triacetyluridine (TAU) or uridine triacetate (PN501), an acyl derivative of uridine such as the acyl derivative described in U.S. Pat. No. 7,582,619 (i.e., 2',3',5'-tri-O-acetonyl uridine), 2,2'-anhydro-5-ethyl uridine, 5- Ethyl-2-deoxyuridine and acyclouridine compounds such as 5-benzyl substituted acyclouridine homologues (including, for example, benzyl acyclouridine, benzyloxybenzyl acyclouridine, aminomethyl-benzyl acyclouridine, aminomethylbenzyloxy-benzyl acyclouridine, hydroxymethyl-benzyloxy-benzyl acyclouridine) (see also, WO89 / 09603 and WO91 / 16315), and dietary supplements derived from sugar cane extracts such as Mitocnol and NucleomaxX.

[0171] UR and its sources include, but are not limited to: meat products, such as fish, pork, beef liver and pancreas, etc.; fungus-related products, such as brewer's yeast, beer, mushrooms, etc.; vegetable products, such as sugar cane, tomatoes, oats, algae, broccoli, etc.; salts, such as phosphate UR, acylated UR, etc. UR and its sources that can be used in the embodiments of the present invention include, but are not limited to, uridine and its sources described in the following U.S. Patents: 9,579,337; 6,316,426 and 5,470,838; the contents related to the compounds disclosed in these publications are incorporated herein by reference.

[0172] UR precursors and their sources include, but are not limited to: prodrugs of UR, such as triphenyluridine, orotic acid, etc.; prodrugs of uridine 5'-monophosphate, such as monoalkyl and dialkyl esters, acyloxyalkyl esters, alkoxycarbonyl methyl esters, substituted ethyl and propyl esters, amide methyl esters, benzyl esters, phenyl esters, phosphoramidates, cyclic phosphates, etc.; UR prodrugs containing monoesters, diesters or triesters of UR, such as monoacetyl, diacetyl and triacetyl UR, etc.; UR prodrugs containing monophosphates, diphosphates or triphosphates of UR, such as monophosphate UR, diphosphate UR, triphosphate UR, etc.; UR homodimers and their esters, such as UPU, etc.; heterodimers of dideoxynucleoside compounds and UR or UPase inhibitors, such as AZT-PU and AZT-P-BAU; and the like. Uridine precursors and their sources that can be used in embodiments of the present invention include, but are not limited to, those described in US Pat. Nos. 5,723,449 and 7,737,128; the compounds disclosed in these publications are incorporated herein by reference.

[0173] Uridine (UR) processing regulator

[0174] If necessary, a UR processing regulator can also be administered to the subject in combination with the UPase inhibitor. UR secretion inhibitory compounds include, but are not limited to, drugs such as delazip and hexobendin. UR secretion inhibitory compounds that can be used in the embodiments of the present invention include, but are not limited to, uridine secretion inhibitory compounds described in the following U.S. Patents: 6,989,376 and 5,567,689; the contents of the compounds disclosed in these publications are incorporated herein by reference.

[0175] UR renal transport competitors include, but are not limited to, drugs such as L-uridine, L-2',3'-dideoxyuridine, D-2',3-dideoxyuridine. UR renal transport competitors that can be used in embodiments of the present invention include, but are not limited to, uridine renal transport competitors described in the following U.S. Patents: 6,989,376; 5,723,449 and 5,567,689; the compounds disclosed in these publications are incorporated herein by reference.

[0176] preparation

[0177] The present invention also provides pharmaceutical compositions used in embodiments of the present invention, for example, pharmaceutical compositions containing UPase inhibitors and / or UR active agents, for example, as described above. The active agent may be present in the pharmaceutical composition in the form of a pharmaceutically acceptable salt, and may be formulated for oral, topical or parenteral administration in the above-mentioned target method. The formulations used in embodiments of the present invention may include a single active agent or a combination of active agents. Therefore, embodiments of the present invention include the following formulations: formulations including a single active agent (e.g., a UPase inhibitor or a UR active agent); and formulations including two or more active agents, wherein the UPase inhibitor and the UR active agent are both present in the same formulation.

[0178] For example, UPase inhibitors and (if necessary) UR active agents (alone or in combination) can be mixed with conventional pharmaceutically acceptable carriers and excipients (e.g., solvents) and used in the form of aqueous solutions, tablets, capsules, elixirs, suspensions, syrups, wafers, etc. In certain embodiments, such pharmaceutical compositions contain about 0.1% to about 90% (by weight) of the active compound, more typically about 1% to about 30% (by weight) of the active compound. The pharmaceutical composition may contain common carriers and excipients, such as corn starch or gelatin, lactose, dextrose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, and alginic acid. Disintegrants commonly used in the formulations of the present invention include cross-linked carboxymethyl cellulose, microcrystalline cellulose, corn starch, sodium starch glycolate, and alginic acid.

[0179] Liquid compositions are generally composed of a suspension or solution of the compound or a pharmaceutically acceptable salt in a suitable liquid carrier, for example, ethanol, glycerol, sorbitol, a non-aqueous solvent such as polyethylene glycol, oil or water, and may contain a suspending agent, a preservative, a surfactant, a wetting agent, a flavoring agent or a coloring agent. Alternatively, liquid preparations may be prepared from reconstituted powders.

[0180] For example, a powder containing the active compound, suspending agent, sucrose and sweetener can be reconstituted with water to form a suspension; and a syrup can be prepared from a powder containing the active ingredient, sucrose and sweetener.

[0181] Compositions in tablet form can be prepared with any suitable pharmaceutical carrier that is commonly used to prepare solid compositions. Examples of such carriers include magnesium stearate, starch, lactose, sucrose, microcrystalline cellulose and binding agents, such as polyvinyl pyrrolidone. Tablets with pigment film coatings or with pigments included as part of the carrier can also be provided. In addition, the active compound can be formulated into a controlled release dosage form, such as a tablet comprising a hydrophilic or hydrophobic matrix.

[0182] The composition in capsule form can be prepared by conventional encapsulation methods, for example, by incorporating the active compound and excipients into hard gelatin capsules. Alternatively, a semisolid matrix of the active compound and high molecular weight polyethylene glycol can be prepared and filled into hard gelatin capsules; or a polyethylene glycol solution of the active compound or a suspension of the active compound in an edible oil (e.g., liquid paraffin or fractionated coconut oil) can be prepared and filled into soft gelatin capsules.

[0183] Tablet binders that may be included are acacia, methylcellulose, sodium carboxymethylcellulose, polyvinyl pyrrolidone (povidone), hydroxypropyl methylcellulose, sucrose, starch and ethylcellulose. Lubricants that may be used include magnesium stearate or other metal stearates, stearic acid, silicone fluids, talc, waxes, oils and silica gel.

[0184] Flavor enhancers may also be used, such as peppermint oil, wintergreen oil, cherry flavor, etc. In addition, it may also be desirable to add a coloring agent to make the dosage form more aesthetically pleasing or to help identify the product.

[0185] For parenteral administration, the compounds of the present invention and their active pharmaceutically acceptable salts may be formulated for intramuscular, intrathoracic or intravenous administration.

[0186] Typical compositions for intramuscular or intrathoracic administration are suspensions or solutions of the active ingredient dissolved in oil (e.g., peanut oil or sesame oil). Typical compositions for intravenous or intrathoracic administration are sterile isotonic aqueous solutions containing a mixture of, for example, active ingredient and dextrose or sodium chloride or dextrose and sodium chloride. Other examples are lactated Ringer's injection, lactated Ringer's solution plus glucose injection, Normosol-M and dextrose, Isolyte E, acylated Ringer's injection, etc. Optionally, cosolvents (e.g., polyethylene glycol), chelating agents (e.g., ethylenediaminetetraacetic acid) and antioxidants (e.g., sodium pyrosulfite) may be included in the formulation. Alternatively, the solution may be lyophilized and then reconstituted with a suitable solvent just before administration.

[0187] The compounds of the present invention and their pharmaceutically acceptable salts which are effective in rectal administration can be formulated into suppositories. Typical suppository formulations usually consist of active ingredients and binders and / or lubricants (such as gelatin or cocoa butter) or other low melting point vegetable or synthetic waxes or fats.

[0188] The compounds of the present invention and pharmaceutically acceptable salts thereof that are effective in topical administration can be formulated into transdermal compositions or transdermal drug delivery devices ("patches"). Such compositions include, for example, substrates, active compound reservoirs, control membranes, linings, and contact adhesives. Such transdermal patches can be used to provide a controlled amount of the compounds of the present invention for continuous or discontinuous infusion. The preparation and use of transdermal patches for delivering drugs are well known in the art. See, for example, U.S. Pat. No. 5,023,252, the contents of which are incorporated herein by reference in their entirety. Such patches can be prepared for continuous, pulsed, or on-demand delivery of drugs.

[0189] In certain embodiments of the invention, the UPase inhibitor and the UR active agent are administered as a single pharmaceutical preparation, which, in addition to the active agent, also includes other suitable compounds and carriers, and can also be used in combination with other active agents. Therefore, the present invention also includes a pharmaceutical composition comprising a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient includes, for example, any suitable solvent, adjuvant, carrier or diluent, and the excipient is easily available to the public. The pharmaceutical composition of the present invention may further contain other active agents known in the art.

[0190] It will be appreciated by those skilled in the art that a variety of suitable methods for administering the preparation of the present invention to a subject or host (e.g., a patient) with corresponding needs are available, and although more than one approach may be used to administer a particular preparation, a particular approach may provide a more rapid and more effective response than another approach. Pharmaceutically acceptable excipients are also well known to those skilled in the art and are readily available. The choice of excipient will depend in part on the specific compound and the specific method used to administer the composition. Therefore, there are numerous suitable formulations of the pharmaceutical composition of the present invention. The following methods and excipients are exemplary methods and reagents only, not restrictive methods and reagents.

[0191] Formulations suitable for oral administration may consist of the following: (a) a liquid solution, such as an effective amount of the compound dissolved in a diluent such as water, saline or orange juice; (b) a capsule, sachet or tablet, each containing a specified amount of the active ingredient as a solid or granules; (c) a suspension suspended in a suitable liquid; and (d) a suitable emulsion. Tablet forms may include one or more of lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, gum arabic, gelatin, colloidal silicon dioxide, carboxymethyl cellulose, talc, magnesium stearate, stearic acid and other excipients, colorants, diluents, buffers, wetting agents, preservatives, flavor enhancers and pharmacologically compatible excipients. Lozenge forms may contain the active ingredient dissolved in flavor enhancers, usually sucrose and acacia or tragacanth; as well as soft pastilles containing the active ingredient in an inert basis such as gelatin and glycerin or sucrose and acacia; emulsions, gels, and the like containing in addition to the active ingredient such excipients as are known in the art.

[0192] The subject formulations of the present invention can be prepared as aerosol formulations for administration by inhalation. These aerosol formulations can be placed in pressurized acceptable propellants, for example, dichlorodifluoromethane, propane, nitrogen, etc. It can also be formulated as a non-pressurized formulation drug, such as a formulation for a nebulizer or sprayer.

[0193] Preparations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, antibacterial agents, and solutes that make the preparation isotonic with the predetermined recipient blood; and aqueous and non-aqueous sterile suspensions, which may include suspending agents, solubilizers, thickeners, stabilizers, and preservatives. The preparations can be packed in unit dose or multi-dose sealed containers (e.g., ampoules and vials), and can be stored under freeze-dried (lyophilized) conditions, and only sterile liquid excipients (e.g., water for injection) need to be added before use. The injections and suspensions prepared at the time of use can be prepared using the aforementioned types of sterile powders, granules, and tablets.

[0194] Formulations suitable for topical administration may be presented as ointments, gels, pastes or foams containing in addition to the active ingredient such other carriers as are known in the art to be appropriate.

[0195] Suppository preparations are also provided by mixing them with various bases (e.g., emulsifying bases or water-soluble bases). Formulations suitable for vaginal administration can be provided as vaginal rings, hemostatic pads, ointments, gels, pastes, foams.

[0196] The present invention can provide unit dosage forms for oral or rectal administration, for example, syrups, elixirs and suspensions, wherein each dosage unit (e.g., a teaspoon, a tablespoon of tablets or suppositories) contains a predetermined amount of a composition containing one or more inhibitors. Similarly, unit dosage forms for injection or intravenous administration can contain the inhibitor in the form of a composition, for example, a solution dissolved in sterile water, saline or other pharmaceutically acceptable carriers.

[0197] The term "unit dosage form" as used herein refers to a physically dispersed single dosage form suitable for human and animal subjects, each unit containing a predetermined amount of the compound described in the present invention, which is mixed with a pharmaceutically acceptable diluent, carrier or solvent in an amount sufficient to produce the desired effect. The specifications of the novel unit dosage form described in the present invention depend on the specific compound used, the effect to be achieved, and the pharmacokinetic characteristics of each compound in the host.

[0198] It will be readily appreciated by those skilled in the art that dosage levels may vary depending on the function of the specific compound, the nature of the delivery vehicle, etc. The appropriate dosage for a given compound can be readily determined by those skilled in the art in a variety of ways.

[0199] In the context of the present invention, the dose applied to an animal, especially a human, should be sufficient to cause a preventive or therapeutic response in the animal within a reasonable time frame. Those skilled in the art recognize that the dose must depend on a variety of factors, including the specification of the specific compound used, the condition of the animal and the weight of the animal, as well as the severity of the disease and the stage of the disease. The size of the dose can also be determined by the presence, nature and extent of any adverse side effects that may accompany the administration of the specific compound. Optionally, the pharmaceutical composition may contain other pharmaceutically acceptable components, such as buffers, surfactants, antioxidants, viscosity modifiers, preservatives, etc. Various such ingredients are well known in the art. For example, see U.S. Pat. No. 5,985,310, the contents of which are incorporated herein by reference.

[0200] Other ingredients suitable for the formulation of the present invention can be found in the following publication: Remington's Pharmaceutical Compendium, Mace Publishing Company, Philadelphia, Pennsylvania, 17th edition (1985). In one embodiment, the cyclodextrin aqueous solution further contains dextrose, such as about 5% dextrose.

[0201] utility

[0202] The targeted methods can be used to treat liver diseases. Although the target liver diseases may vary, in some cases, the liver diseases are characterized by the presence of characteristic fibrosis; or accumulation of extracellular matrix molecules that constitute scar tissue as a toxic endpoint; and other diseases, such as, among others, pulmonary fibrosis, renal fibrosis, systemic sclerosis (SS), scleroderma graft-versus-host disease, radiation-induced lung fibrosis, and cardiac fibrosis.

[0203] In some cases, the liver disease is fatty liver disease. Fatty liver disease, also referred to as fatty liver or fatty liver disease (FLD), refers to the disease of accumulating a large amount of triglycerides in liver cells by steatosis or abnormal retention of intracellular lipids. Although there are multiple causes, fatty liver is still regarded as a single disease, common in excessive drinking and obese subjects (with or without the influence of insulin resistance). This disease is also related to other diseases that affect fat metabolism. FLD can be divided into two different disease types: alcoholic FLD and non-alcoholic FLD. Both diseases show microvesicular and macrovesicular steatosis in different stages of the disease. The accumulation of fat may also be accompanied by progressive liver inflammation (hepatitis), called steatohepatitis. Fatty liver is also referred to as alcoholic steatosis and non-alcoholic fatty liver disease (NAFLD) in the art, and more serious forms are alcoholic steatohepatitis (belonging to alcoholic liver disease) and non-alcoholic steatohepatitis (NASH). Nonalcoholic fatty liver disease-related cirrhosis is the most severe form of the disease and is characterized by liver inflammation, which can lead to scarring of liver tissue and ultimately liver failure. In some cases, the liver disorder is NAFLD, NASH, or DILI.

[0204] "Treatment" refers to at least alleviation of the symptoms associated with the condition afflicting the host, wherein alleviation, used in a broad sense, refers to at least a reduction in the degree of a parameter (e.g., a symptom associated with the condition being treated or a side effect caused by the administration of a drug). Thus, treatment also includes situations in which the pathological condition or at least the symptoms associated with the condition are completely suppressed, such as prevented from occurring, or stopped (e.g., terminated) so that the host is no longer afflicted with the condition or at least no longer experiences the symptoms characteristic of the condition. Treatment also includes prophylactic treatment of a subject so that the subject does not develop a liver condition. Thus, treatment includes preventing a subject from developing a liver condition.

[0205] A plurality of patients may be treated according to the subject methods. Typically, such a host is a "mammal," a term broadly used to describe organisms belonging to the class mammals, including carnivores (e.g., dogs and cats), rodents (e.g., mice, guinea pigs, and rats), and primates (e.g., humans, chimpanzees, and monkeys). In many embodiments, the subject is a human.

[0206] In certain embodiments, the subject is a subject who has been diagnosed with a disease, and thus needs to administer the active agent. In certain embodiments, the method may include diagnosing whether the subject is suffering from a disease to be treated by administering the active agent.

[0207] When the liver disease is DILI, the method of the present invention can be used in combination with a treatment regimen suspected of causing DILI, such as treating DILI that has already occurred or treating DILI prophylactically. When the compounds of the present invention are administered in combination with other therapies, the dosage of the compound used in combination will inevitably vary depending on the type of compound drug used, the specific drug used, the condition treated, etc. The terms "combination therapy", "combination therapy" or "combination therapy" as used herein are used interchangeably and refer to treating an individual with at least two different therapeutic agents. The terms "combination" or "combination administration" and the like as used herein are intended to cover the administration of selected therapeutic agents to a single patient, and are intended to include treatment regimens in which the agents are not necessarily administered by the same route or at the same time. The term "drug combination" refers to a product obtained by mixing or combining more than one active ingredient, including fixed and non-fixed combinations of active ingredients. "Fixed combination" means that the active ingredients (e.g., the compounds disclosed herein and one or more additional therapeutic agents) are all administered to the patient simultaneously in the form of a single entity or dosage. "Non-fixed combination" means that the active ingredients (e.g., a compound disclosed herein and one or more additional therapeutic agents) are administered to a patient simultaneously, simultaneously or sequentially (without specific time limits) in the form of a single entity, wherein such administration is such that the two compounds achieve therapeutically effective levels in the patient's body. The latter also applies to cocktail therapy, such as administration of 3 or more active ingredients. As used herein, the methods of the invention can be used in combination with one or more additional therapeutic agents, such as, but not limited to, therapeutic agents for pulmonary arterial hypertension, such as ambrisentan, bosentan, treprostinil, sildenafil, epoprostenol, treprostinil, iloprost, aldosterone receptor antagonists such as spironolactone and eplerenone; angiotensin converting enzyme inhibitors, such as trandolapril, fosinopril, enalapril, captopril, ramipril, moexipril, lisinopril, quinapril, benazepril, and perindopril; angiotensin II inhibitors, such as eprosartan, olmesmian, telmismian, losartan, valsmian, candesartan, and irbesmian; antianginal drugs, such as nitrates; triglyceride, isosorbide mononitrate and isosorbide dinitrate; antiarrhythmics, including moricizine, quinidine, disopyramide, phenytoin, propafenone, flecamide, mexiletine, lidocaine, procainamide, propranolol, acebutolol, amiodarone, dofetilide, dronedarone, sotalol, ibutilide, diltiazem, verapamil, nifedipine, nimodipine, felodipine, nicardipine, clevidipine, isradipine, bepridil, nisoldipine, adenosine, and digoxin; beta-adrenergic receptor antagonists, such as betaxolol, bisoprolol, metoprolol, atenolol, nebivolol, nadolol, carvedilol, labetalol, timolol, carteolol, penbutolol, pindolol, and esmolol;Antidiabetic agents, including secretagogues such as sulfonylureas, tolbutamide, acetohexamide, tolazamide, chlorpropamide, glipizide, glyburide, glimepiride, glyburide, gliclazide, meglitinides such as nateglinide, senaglinide, repaglinide, insulin sensitizers such as biguanides, metformin, thiazolidinediones such as rosiglitazone, isaglinide, darglitazone, englitazone and pioglitazone; alpha-glucosidase inhibitors such as miglitazone, voglib glucagon-like peptide analogs and agonists, such as exenatide, liraglutide, taspglutide, dipeptidyl peptidase-4 inhibitors such as vildagliptin, sitagliptin, and saxagliptin; amylin analogs, such as pramlintide; ligands or agonists of peroxisome proliferator-activated receptor (PPAR)-α, β, δ, and γ cholesterol-lowering agents, such as hydroxymethylglutaryl-coenzyme A (HMG-CoA ) reductase inhibitors like statins, for example, atorvastatin, chlorvastatin, lovastatin, pitavastatin, pravastatin, rosovastatin and simvastatin; agonists of the vitamin A class X receptor (RXR), for example, ALRT-268, LG-1268 or LG-1069; glucokinase activators, inhibitors of liver enzymes involved in the stimulation of gluconeogenesis and / or glycogenolysis; diuretics, for example, acetazolamide, dichlorphenamide, methylphenidate azolamide, torsemide, furosemide, bumetanide, ethacrynic acid, amiloride, triaminopterin, indapamide, metolazone, methylclothiazide, hydrochlorothiazide, chlorothiazide, metolazone, bendrofluazide, polythiazide, and chlorthalidone; vasodilators such as alprostadil, hydralazine, minoxidil, nesiritide, and nitroprusside; lipid-lowering drugs such as cholestyramine, colestipol, clofibrate, gemfibrozil, procobuc, or dextrothyroxine. ;

[0208] Kits and Systems

[0209] The present invention also provides kits and systems that can be used to implement methods such as those described above. For example, kits and systems for implementing the subject methods may include one or more pharmaceutical preparations, such preparations including UPase inhibitors, and in some embodiments, they include UR active agents. Therefore, in certain embodiments, the kit may include a single pharmaceutical composition in one or more unit dosage forms, wherein the composition includes a UPase inhibitor and a UR active agent. In other embodiments, the kit may include two or more separate pharmaceutical compositions, each containing a UPase inhibitor and optionally a UR active agent.

[0210] In addition to the above components, the target kit may further include instructions for implementing the target method. These instructions may be present in the target kit in a variety of forms, one or more of which may be present in the kit. One form in which these instructions may exist is printed information printed on a suitable medium or substrate (e.g., a sheet of paper or multiple sheets of paper with information printed thereon), a kit package, a package insert, etc. Another form in which it exists is a computer-readable medium on which information has been recorded, such as a floppy disk, a CD, etc. Another form in which these instructions may exist is a website, which can be used to access information on a remote website via the Internet. Any suitable device may be present in the kit. For example, a kit according to one embodiment includes a first component (a) instructions for use of a plasma UR level regulator; and a second component (b) a pharmaceutical composition comprising uridine, a UR prodrug or a UR mimetic.

[0211] A specific kit of interest is a kit comprising a 2,2'-anhydropyrimidine pharmaceutical composition of the invention and suitable for practicing the subject method of the invention, for example, suitable for alleviating severe liver disorders.

[0212] The term "system" as used herein refers to a collection of UPase inhibitors and optional UR active agents present in a single or different composition, which are combined together to implement the subject method. For example, separately obtained UPase inhibitors and UR active agent dosage forms combined together and co-administered to a subject according to the present invention are systems according to the present invention.

[0213] The following examples further illustrate the present invention but should not be construed as limiting the scope of the present invention in any way.

[0214] Example

[0215] I.UR increases with the increase of compound I concentration.

[0216] Since UR is cleared quickly, it can eliminate t 1 / 2 Just a few minutes, 46 The elimination of compound 1 in mice 1 / 2 The plasma UR concentration is measured after administration of discrete doses of Compound I (e.g., for ip administration) as the plasma UR concentration is only 1-2 hours. Therefore, BDF-1♂ mice were continuously infused with Compound I (genuine TK-112690, batch number TCY90108) via a subcutaneously implanted osmotic pump and plasma UR concentrations were measured.

[0217] Prepare a solution of Compound I at a concentration of 500 mg / mL in sterile PBS. Osmotic pump ( Mini-osmotic pumps (2001D and 1003D, Alza Co) were loaded with 200 μL (2001D osmotic pump) and / or 100 μL (1003D osmotic pump) of TK-112690 solution.

[0218] BDF-1 male mice (n=6) were treated with Compound I at a dose of 667, 833, or 3000 mg / kg / day via subcutaneously implanted osmotic pumps at a constant rate. Animals were anesthetized with 100 mg / kg ketamine prior to pump implantation. An approximately 1 cm incision was made on the dorsal surface near the animal's scapula using surgical scissors. A subcutaneous tunnel was created in the anterior end of the animal using hemostats. The osmotic pump was placed in the subcutaneous tunnel. The incision was sealed with wound clips.

[0219] Blood was collected from animals anesthetized with ketamine (ip 100 mg / kg). Blood samples were collected from animals receiving constant-speed infusion of TK-112690 at 72 hours (667 mg / kg / day and 833 mg / kg / day) and 24 hours (3000 mg / kg / day) after pump implantation. Whole blood (~0.8 mL) was drawn through the retroorbital sinus using a heparin-coated microhematocrit tube and collected in an EDTA microtube. The blood sample was transferred to a new 1.5 mL microcentrifuge tube and centrifuged at a centrifugal force of 14,000 x g for 10 minutes using an Eppendorf Minispin Plus stored in a 4°C refrigerator. Accurately transfer 0.4 mL of plasma to a new microcentrifuge tube containing 2 μL of 10 mM 5-FU and vortex centrifuge for about 5 seconds at the highest setting. 5-FU with a final concentration of 50 μM was used as an internal standard. Animals were sacrificed by cervical dislocation and disposed of appropriately.

[0220] Blood samples were collected from animals receiving constant rate infusion of Compound I at 72 hours (667 mg / kg / day and 833 mg / kg / day) and 24 hours (3000 mg / kg / day) after pump implantation. Whole blood (~0.8 mL) was drawn through the retro-orbital sinus using a heparin-coated microhematocrit tube and collected in an EDTA microtube. The blood sample was transferred to a new 1.5 mL microcentrifuge tube and centrifuged at a centrifugal force of 14,000 x g for 10 minutes using an Eppendorf Minispin Plus stored in a 4°C refrigerator. Accurately transfer 0.4 mL of plasma to a new microcentrifuge tube containing 2 μL of 10 mM 5-FU and vortex centrifuge for about 5 seconds at the highest setting. 5-FU at a final concentration of 50 μM was used as an internal standard. The animals were killed by cervical dislocation and disposed of appropriately.

[0221] Solid phase extraction (SPE) was performed on the analytes (UR, Compound I, and 5-FU) in plasma prior to HPLC analysis. Supelco C8 SPE columns were used for the extraction process. All solutions were pushed through the SPE columns using positive pressure generated by a vacuum pressure pump (Barnant Company Model 400-1901). The flow rate through the SPE column was approximately 2 drops per second. The SPE column was pre-washed with a total of 2.4 mL of sterile PBS (room temperature; pH = 7.4). 0.6 mL of PBS was added to the SPE column in four portions and pushed through the extraction column. After pre-washing, all (0.4 mL) of the plasma sample (with the addition of 5-FU internal standard) was immediately transferred to the extraction column and pushed through the extraction column. 0.5 mL of 5M NaCl (room temperature; pH ~ 5) was accurately pushed through the SPE column to separate the analytes from the SPE column. The eluted sample was collected in a new 1.5 mL microcentrifuge tube. The sample was transferred to a new HPLC vial and analyzed.

[0222]

[0223] HPLC analysis was performed at room temperature (RT) using a ThermoFinnigan Spectra system equipped with a degasser, pump, autosampler, and UV detector. Chromatograms were constructed using a chart recorder equipped with a pen. Analytes were separated using a Phenomenex C18 reverse phase column (250x4.6mm). Two separate mobile phase gradients were used for HPLC analysis: (1) 5% methanol in nanowater containing 0.1% formic acid (2) 5% methanol in acetonitrile containing 0.1% formic acid (flow rate = 0.5mL / min). The HPLC responses of Compound I and UR were divided by the 5-FU response. These ratios were converted to concentrations of Compound 1 using a calibration curve.

[0224] For regression analysis of the study data (UR concentration versus compound 1 concentration), see Figure 1 Higher concentrations of compound 1 were seen to be associated with higher levels of UR.

[0225] II. Methionine-choline diet (MCD) model of NASH.

[0226] Mice fed an MCD diet are a standard model of diet-induced NASH. 47,48All animals were housed in well-ventilated standard cages throughout the experimental period. All animals had free access to tap water. Eight-week-old male C57BL / 6 mice from Charles River were acclimated, fed with standard diet for 3 days, and group-housed in hepa-filtered cages (5 animals per cage) with a normal 12-hour light cycle (lights off from 8 a.m. to 8 p.m.). Temperature and humidity were 22±2°C and 50±10%, respectively. Cages were changed once a week.

[0227] After acclimation, animals were randomized into similar treatment groups according to body weight and allowed to consume a diet deficient in methionine and choline ad libitum: 4.2 kcal / g; MP Biomedicals, Solon, OH. Food and water intake was measured 3 times per week (concurrently with body weight measurements). After the acclimation period, 24 mice (n = 6 / treatment group) were weighed 3 times per week until sacrifice.

[0228] The 4 treatment groups were as follows:

[0229] ·Group 1: solvent+MCD

[0230] Group 2: MCD + 200 mg / kg UR

[0231] Group 3: MCD + 60 mg / kg Compound I

[0232] Group 4: MCD + 60 mg / kg Compound 1 plus 200 mg / kg UR (administered 30 minutes after Compound 1)

[0233] Compound 1 and UR (unit: mg / kg, dissolved in 10 mL / kg vehicle) were injected intraperitoneally (ip) twice daily (bid) with an interval of at least 8 hours for 28 days. Vehicle = PBS. Vehicle, UR and compound 1 ± UR were given once daily starting 2 days before the animals received the MCD diet. Mice received the MCD diet for 26 days.

[0234] After 26 days of MCD diet, animals were sacrificed approximately 2 hours after the last dose and non-fasting blood glucose and lipid profiles, TNF-α, ALT, and AST were measured. Total body weight was measured once a week. The total liver was weighed, the inner lobe was removed, fixed with formalin, and stained with Oil Red O to assess lipid content. The intensity score of fixed tissue sections measured by an independent histopathologist was also determined.

[0235] As expected, 49 All treatment groups lost approximately 20% of their body weight ( Figure 2 , data are expressed as mean + / - SEM). There was no difference in body weight among the groups.

[0236] There were also no differences between groups 2, 3, and 4 with respect to total triglycerides, cholesterol, AST, or ALT. For HDL-cholesterol values, there was a statistically significant difference between group 1 and group 4 ( Figure 3 ) (Serum HDL cholesterol concentrations measured at the end of the MCD study. Data are mean + / - SEM. TK-90 is compound 1, also known as TK-112690). Most importantly, the finding that fibrosis occurred ( Figure 4 )((A) Representative H&E images of liver sections from each experimental group. (B) Fibrosis scores for the images shown in (A). TK-90 is compound 1, also known as TK-112690. Data are mean + / - SEM.) This data is important because fibrosis is a pathological endpoint of NASH. Figure 4 According to the data in , there were statistically significant differences between groups 2, 3 and 4 and group 1 (control group).

[0237] Although the claims are appended hereto, the scope of the present invention is further defined by the following clauses:

[0238] 1. A method of treating a liver disorder in a subject, the method comprising:

[0239] An effective amount of 2,2'-anhydropyrimidine or a derivative thereof is administered to the subject to treat the liver disorder in the subject.

[0240] 2. The method according to clause 1, wherein the 2,2'-anhydropyrimidine or its derivative is a compound represented by formula (I):

[0241] (I)

[0242] or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof;

[0243] in:

[0244] Each R 1 , R 2 , R 3 and R 4 are independently selected from the group consisting of hydrogen, substituted or unsubstituted heteroatoms, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, carbohydrates, nucleic acids, amino acids, peptides, dyes, fluorophores and polypeptides.

[0245] 3. The method according to clause 2, wherein each R 1 , R 2 , R3 and R 4 Each is independently selected from the group consisting of hydrogen, hydroxyl, mercapto, amino, hydroxymethyl, methoxy, halogen, pseudohalogen and substituted or unsubstituted lower hydrocarbon containing 1 to 20 carbon atoms.

[0246] 4. The method according to clause 2, wherein the lower hydrocarbon is selected from the group consisting of alkyl, alkenyl, alkanoyl, aryl, aroyl, aralkyl, alkylamino and esters thereof.

[0247] 5. The method according to clause 2, wherein R 1 is hydrogen, fluorine, methyl, ethyl, propyl, benzyl or 2-bromovinyl; R 2 is hydrogen, hydroxyfluoride, methyl, ethyl, propyl, benzyl, benzoyl, benzoyloxy or 2-bromovinyl; and each R 3 and R 4 are independently selected from the group consisting of hydroxyl and benzoyloxy.

[0248]

[0249] 6. The method according to clause 5, wherein R 1 is hydrogen or methyl; R 2 is hydrogen; and each R 3 and R 4 are independently selected from the group consisting of hydroxyl and benzoyloxy.

[0250] 7. The method according to clause 1, wherein the 2,2'-anhydropyrimidine or a derivative thereof is selected from the group consisting of: 2,2'-anhydro-5-methyluridine; 3'-O-benzoyl-2,2'-anhydrouridine; 3'-O-benzoyl-2,2'-anhydro-5-methyluridine; 5'-O-benzoyl-2,2'-anhydrouridine and 5'-O-benzoyl-2,2'-anhydro-5-methyluridine.

[0251] 8. The method according to clause 7, wherein the 2,2'-anhydropyrimidine or a derivative thereof is 2,2'-anhydro-5-methyluridine.

[0252] 9. The method according to clause 7, wherein the 2,2'-anhydropyrimidine or a derivative thereof is 3'-O-benzoyl-2,2'-anhydro-5-methyluridine.

[0253] 10. The method according to clause 7, wherein the 2,2'-anhydropyrimidine or a derivative thereof is 5'-O-benzoyl-2,2'-anhydro-5-methyluridine.

[0254] 11. The method according to clause 1, wherein the 2,2'-anhydropyrimidine or a derivative thereof comprises stereoisomers.

[0255] 12. The method according to clause 11, wherein the stereoisomer is selected from the group consisting of 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil, 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-uracil, 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil, 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-uracil and 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil.

[0256] 13. The method according to any of the preceding clauses, wherein the liver disorder is selected from the group consisting of NAFLD, NASH and DILI.

[0257] 14. A system according to any of the preceding clauses, wherein the treatment is a prophylactic treatment.

[0258] 15. The method according to clause 14, wherein the liver disorder is DILI.

[0259] 16. The method according to any one of clauses 1 to 13, wherein the subject suffers from the liver disorder.

[0260] 17. A method of treating a liver disorder in a subject, the method comprising:

[0261] An effective amount of 2,2'-anhydropyrimidine or a derivative thereof and a uridine (UR) active agent is co-administered to the subject to treat the liver disorder in the subject.

[0262] 18. The method according to clause 17, wherein the 2,2'-anhydropyrimidine or its derivative is a compound represented by formula (I):

[0263] (I)

[0264] or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof;

[0265] in:

[0266] Each R 1 , R 2 , R 3 and R 4are independently selected from the group consisting of hydrogen, substituted or unsubstituted heteroatoms, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, carbohydrates, nucleic acids, amino acids, peptides, dyes, fluorophores and polypeptides.

[0267] 19. The method according to clause 18, wherein each R 1 , R 2 , R 3 and R 4 Each is independently selected from the group consisting of hydrogen, hydroxyl, mercapto, amino, hydroxymethyl, methoxy, halogen, pseudohalogen and substituted or unsubstituted lower hydrocarbon containing 1 to 20 carbon atoms.

[0268] 20. The method according to clause 18, wherein the lower hydrocarbon is selected from the group consisting of alkyl, alkenyl, alkanoyl, aryl, aroyl, aralkyl, alkylamino and esters thereof.

[0269] 21. The method according to clause 18, wherein R 1 is hydrogen, fluorine, methyl, ethyl, propyl, benzyl or 2-bromovinyl; R 2 is hydrogen, hydroxyfluoride, methyl, ethyl, propyl, benzyl, benzoyl, benzoyloxy or 2-bromovinyl; and each R 3 and R 4 are independently selected from the group consisting of hydroxyl and benzoyloxy.

[0270] 22. The method according to clause 21, wherein R 1 is hydrogen or methyl; R 2 is hydrogen; and each R 3 and R 4 are independently selected from the group consisting of hydroxyl and benzoyloxy.

[0271] 23. A method according to clause 18, wherein the 2,2'-anhydropyrimidine or a derivative thereof is selected from the group consisting of: 2,2'-anhydro-5-methyluridine; 3'-O-benzoyl-2,2'-anhydrouridine; 3'-O-benzoyl-2,2'-anhydro-5-methyluridine; 5'-O-benzoyl-2,2'-anhydrouridine and 5'-O-benzoyl-2,2'-anhydro-5-methyluridine.

[0272] 24. The method according to clause 23, wherein the 2,2'-anhydropyrimidine or a derivative thereof is 2,2'-anhydro-5-methyluridine.

[0273] 25. The method according to clause 23, wherein the 2,2'-anhydropyrimidine or a derivative thereof is 3'-O-benzoyl-2,2'-anhydro-5-methyluridine.

[0274] 26. The method according to clause 23, wherein the 2,2'-anhydropyrimidine or a derivative thereof is 5'-O-benzoyl-2,2'-anhydro-5-methyluridine.

[0275] 27. The method according to clause 18, wherein the 2,2'-anhydropyrimidine or a derivative thereof comprises stereoisomers.

[0276] 28. A method according to clause 27, wherein the stereoisomer is selected from the group consisting of 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil, 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-uracil, 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil, 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-uracil and 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil.

[0277] 29. The method according to any one of clauses 17 to 28, wherein the liver disorder is selected from the group consisting of NAFLD, NASH and DILI.

[0278] 30. The method according to any one of clauses 17 to 29, wherein the treatment is prophylactic treatment.

[0279] 31. The method according to clause 30, wherein the liver disorder is DILI.

[0280] 32. The method according to any one of clauses 17 to 29, wherein the subject suffers from the liver disorder.

[0281] In at least some of the foregoing embodiments, one or more elements used in one embodiment may be used interchangeably in another embodiment, unless such replacement is technically infeasible. It will be appreciated by those skilled in the art that various other omissions, additions, and modifications may be made to the above methods and structures without departing from the claimed subject matter. All of these modifications and changes are intended to be covered by the subject matter scope defined in the appended claims.

[0282] Those skilled in the art will understand that, in general, the terms used herein, particularly in the appended claims (e.g., in the body of the appended claims), should generally be understood as "open" terms (e.g., the term "including" should be interpreted as "including but not limited to", the term "having" should be interpreted as "having at least", etc.). Those skilled in the art will also understand that if a specific number is intended to be indicated in an introduced claim, such intention will be explicitly indicated in the claim, and in the absence of such explicit indication, such intention will be deemed not to exist. For example, to aid understanding, the appended claims may use the introductory phrases "at least one" and "one or more" to introduce features in the claims. However, the use of such phrases should not be interpreted as implying that a claim feature introduced by the indefinite article "a" or "an" limits any particular claim containing that feature to embodiments containing only one of that feature, even if the claim includes both the introductory phrase "one or more" or "at least one" and an indefinite article such as "a" or "an" (e.g., "a" and / or "an" should be interpreted as meaning "at least one" or "one or more"); the same is true when a definite article is used to introduce a feature in a claim. In addition, even if a specific number of introduced claim features is explicitly stated, those skilled in the art will recognize that such enumeration should be interpreted as meaning at least the listed number (e.g., the phrase "two features" without other modifiers means at least two of the features, or two or more of the features). In addition, when using expressions such as "at least one of A, B, and C, etc.", it should generally be interpreted according to the meaning of the expression generally understood by those skilled in the art (for example, "a system having at least one of A, B, and C" should include but not be limited to systems having A alone, B alone, C alone, A and B, A and C, B and C, and / or A, B, and C, etc.). When using expressions such as "at least one of A, B, or C, etc.", it should generally be interpreted according to the meaning of the expression generally understood by those skilled in the art (for example, "a system having at least one of A, B, or C" should include but not be limited to systems having A alone, B alone, C alone, A and B, A and C, B and C, and / or A, B, and C, etc.). Those skilled in the art should also understand that any transitional conjunctions and / or phrases that substantially represent two or more optional items, whether in the specification, claims, or drawings, should be understood to give the possibility of including one of these items, either of these items, or both of these items. For example, the phrase "A or B" should be understood to include the possibility of "A" or "B" or "A and B".

[0283] In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0284] It will be understood by those skilled in the art that for any and all purposes, such as to provide a written description, all ranges disclosed herein also include any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily regarded as fully describing and achieving at least bisection, trisection, quartering, quintillation, decimalization, etc. of the range. As a non-limiting example, each range discussed herein can be easily divided into the lower third, the middle third, and the upper third, etc. It will be understood by those skilled in the art that all languages ​​such as "until", "at least", "greater than", "less than" and the like include the listed numbers and refer to the ranges that can be subsequently divided into sub-ranges as described above. Finally, it will be understood by those skilled in the art that the range includes each individual number. Therefore, for example, a group having 1-3 units refers to a group having 1, 2 or 3 units. Similarly, a group having 1-5 units refers to a group having 1, 2, 3, 4 or 5 units, and so on.

[0285] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, in view of the teachings of the invention, it will be apparent to one skilled in the art that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0286] Therefore, the foregoing only illustrates the principles of the present invention. It should be understood that those skilled in the art can design various structures, although not explicitly stated or shown here, these designs reflect the principles of the present invention and do not exceed the spirit and scope of the present invention. In addition, all examples and conditional language listed herein are mainly to help readers understand the principles of the present invention and the ideas provided by the inventors to further expand the art, and should be interpreted as not being limited by these specific examples and conditions. Moreover, all statements citing the principles, aspects and embodiments of the present invention and their specific examples herein are intended to cover their equivalents in structure and function. In addition, the equivalents are intended to include currently known equivalents and equivalents to be developed in the future, that is, any functionally identical elements developed without considering their structure. Moreover, regardless of whether the disclosure of the present invention is clearly described in the claims, any content thereof will not be disclosed to the public.

[0287] Therefore, the scope of the present invention is not limited to the exemplary embodiments shown and described herein. Instead, the scope and spirit of the present invention are embodied by the appended claims. In the claims, only when the phrase "means for..." or "step for..." is explicitly used at the beginning of the limitation content of the claim, "35 U.S.C. Section 112(f)" or "35 U.S.C. Section 112(6)" is explicitly defined as being cited; if the phrase is not used in the limitation content of the claim, "35 U.S.C. Section 112(f)" or "35 U.S.C. Section 112(6)" is not cited.

Claims

1. A method of treating a liver disorder in a subject, the method comprising: An effective amount of 2,2'-anhydropyrimidine or a derivative thereof is administered to the subject to treat the liver disorder in the subject.

2. The method according to claim 1, wherein the 2,2'-anhydropyrimidine or its derivative is a compound represented by formula (I): (I) or a pharmaceutically acceptable salt, solvate, hydrate, prodrug form thereof and stereoisomers thereof; in: Each R 1 , R 2 , R 3 and R 4 are independently selected from the group consisting of hydrogen, substituted or unsubstituted heteroatoms, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aralkyl, carbohydrates, nucleic acids, amino acids, peptides, dyes, fluorophores and polypeptides.

3. The method according to claim 2, wherein each R 1 , R 2 , R 3 and R 4 Each is independently selected from the group consisting of hydrogen, hydroxyl, mercapto, amino, hydroxymethyl, methoxy, halogen, pseudohalogen and substituted or unsubstituted lower hydrocarbon containing 1 to 20 carbon atoms.

4. The method according to claim 2, wherein the lower hydrocarbon is selected from the group consisting of alkyl, alkenyl, alkanoyl, aryl, aroyl, aralkyl, alkylamino and esters thereof.

5. The method according to claim 2, wherein R 1 is hydrogen, fluorine, methyl, ethyl, propyl, benzyl or 2-bromovinyl; R 2 is hydrogen, hydroxyfluoride, methyl, ethyl, propyl, benzyl, benzoyl, benzoyloxy or 2-bromovinyl; and each R 3 and R 4 are independently selected from the group consisting of hydroxyl and benzoyloxy.

6. The method according to claim 5, wherein R 1 is hydrogen or methyl; R 2 is hydrogen; and each R 3 and R 4 are independently selected from the group consisting of hydroxyl and benzoyloxy.

7. The method of claim 1, wherein the 2,2'-anhydropyrimidine or a derivative thereof is selected from the group consisting of: 2,2'-anhydro-5-methyluridine; 3'-O-benzoyl-2,2'-anhydrouridine; 3'-O-benzoyl-2,2'-anhydro-5-methyluridine; 5'-O-benzoyl-2,2'-anhydrouridine and 5'-O-benzoyl-2,2'-anhydro-5-methyluridine. The method according to claim 1 , wherein the 2,2′-anhydropyrimidine or a derivative thereof comprises stereoisomers.

9. The method of claim 8, wherein the stereoisomer is selected from the group consisting of 2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil, 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-uracil, 3'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil, 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-uracil and 5'-O-benzoyl-2,2'-anhydro-1-(β-D-arabinofuranosyl)-5-methyluracil.

10. The method according to any one of the preceding claims, wherein the liver disorder is selected from the group consisting of NAFLD, NASH and DILI.

11. The system according to any one of the preceding claims, wherein the treatment is a prophylactic treatment.

12. The method of claim 14, wherein the liver disorder is DILI.

13. The method of any one of claims 1 to 10, wherein the subject suffers from the liver disorder.

14. The method of any one of the preceding claims, wherein the method comprises administering to the subject an effective amount of a 2,2'-anhydropyrimidine or a derivative thereof and a uridine (UR) active agent in combination to treat the liver disorder in the subject.

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