CN114053293A

CN114053293A - Use of heterocyclic compounds for the treatment or prophylaxis of viral hepatitis

Info

Publication number: CN114053293A
Application number: CN202011594937.8A
Authority: CN
Inventors: 李瑛颖; 陈明键; 仇思念
Original assignee: China Israel Hyde Artificial Intelligence Drug Research And Development Co ltd
Current assignee: China Israel Hyde Artificial Intelligence Drug Research And Development Co ltd
Priority date: 2020-12-29
Filing date: 2020-12-29
Publication date: 2022-02-18

Abstract

The invention provides an application of heterocyclic compound in preparing a medicament for treating or preventing viral hepatitis. The compound is a compound of a general formula 1 or a general formula 2 or a pharmaceutically acceptable salt thereof, and preferably, the viral hepatitis is hepatitis B. The invention also provides a pharmaceutical composition for treating or preventing viral hepatitis comprising a compound of formula 1 or a pharmaceutically acceptable salt thereof, optionally one or more additional therapeutic or prophylactic agents, and a pharmaceutically acceptable carrier.

Description

Use of heterocyclic compounds for the treatment or prophylaxis of viral hepatitis

Technical Field

The invention relates to the technical field of antiviral drugs, in particular to a pharmaceutical composition for treating or preventing viral hepatitis and application thereof.

Background

Human Hepatitis B Virus (HBV) infection is a major public health problem worldwide. After acute hepatitis B virus infection, about 8% of hepatitis B virus still develops into chronic hepatitis B infection, and persistent HBV infection can cause cirrhosis and even liver cancer. China is the big country of hepatitis B, and hepatitis B virus carriers are close to 1.3 hundred million people and account for about 9 percent of the total population. Although the new hepatitis B infection rate is effectively controlled along with the wide popularization of hepatitis B vaccines, the population base of hepatitis B virus carriers is large, and the prevention and treatment of hepatitis B become the central importance of public health problems in China. The hepatitis B transmission pathway is mainly through vertical transmission and horizontal transmission. Vertical transmission refers to mother-to-baby transmission; horizontal transmission is primarily through the blood.

The treatment of hepatitis B is also a long-term process, and the treatment aims to inhibit or eliminate HBV to the maximum extent, relieve inflammation and necrosis of liver cells and liver fibrosis, delay and stop the progress of diseases, reduce and prevent liver decompensation, liver cirrhosis, hepatocellular carcinoma and complications thereof, thereby improving the quality of life and prolonging the survival time.

There are many hepatitis b therapeutic drugs on the market today, mainly by antiviral treatment with interferon or nucleoside analogues. In the case of interferon, recombinant DNA leukocyte interferon (IFN-. alpha.) inhibits the replication of HBV. However, when the interferon is used for treating hepatitis B, strong adverse reactions are often accompanied, including bone marrow suppression, thyroid function influence, depression and the like.

Nucleoside analogues inhibit HBV production primarily by inhibiting reverse transcriptase activity during HBV replication, and clinically useful drugs include the following classes: lamivudine and famciclovir, such as acyclovir, adefovir, entecavir, tenofovir, foscarnet and the like, and the medicaments have certain HBV inhibiting effect.

Although these reverse transcriptase inhibitors can effectively reduce HBV DNA level and make patients control HBV level, they have no direct effect on the clearance of HBV cccDNA and HBsAg because their target of action is the process of RNA reverse transcription into DNA. Therefore, the seroconversion probability of HBsAg is very low in single-drug treatment of nucleoside analogue, hepatitis B cannot be really cured, and patients need to take the drug for a long time or even for life.

Under the condition of taking the above drugs for a long time, the problems of drug resistance, huge medical cost, serious side effects of the drugs and the like are a heavy burden for hepatitis B patients. The key point is that at present, no medicine can completely eliminate viruses to achieve the functional cure of hepatitis B. Therefore, the urgent need in the art is to provide a new drug for treating hepatitis b, which can eliminate HBsAg and achieve a functional cure.

Disclosure of Invention

According to the invention, through an artificial intelligence system, based on a plurality of hepatitis B treatment target spots and big data analysis, the compound of the general formula 1 with the hepatitis B treatment effect is screened out, and further through verification of biological experiments, the compound of the general formula 1 with the effect of removing HBsAg and HBeAg is obtained, so that the hepatitis B can be cured functionally and removed.

In one embodiment, the present invention provides the use of a compound of formula 1 or formula 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prevention of viral hepatitis,

in the general formula 1 and the general formula 2, R₁And R is at least one selected from deuterium, halogen, amino, hydroxyl, C1-3 alkoxy, C6-12 aryloxy, amido, substituted or unsubstituted C1-3 alkanoyloxy, C6-C12 aroyloxy, C1-10 carboxyl and substituted or unsubstituted C1-C10 alkyl respectively,

q, X, Y and Z are each selected from C, N, O and S,

the substituent of the substituted C1-10 alkyl is selected from at least one of deuterium, halogen, hydroxyl, amino, amido, C1-3 alkanoyloxy and C1-10 carboxyl,

the substituent of the substituted C1-3 alkanoyloxy group is selected from deuterium and a C6-C12 aryl group,

m is an integer of 1 to 2, n is an integer of 1 to 3,

preferably, the pharmaceutically acceptable salt is selected from at least one of the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, thiocyanate, toluenesulfonate, undecanoate, sodium salt, calcium salt, potassium salt, ammonium salt, tetraethylammonium salt, methylammonium salt, dimethylammonium salt, and ethanolamine salt.

In a preferred embodiment, the present invention provides the use of a compound of formula 1 or formula 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for reducing or eliminating HBsAg and/or HBeAg.

Such pharmaceutically acceptable salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, malate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, thiocyanate, toluenesulfonate, undecanoate, sodium salt, calcium salt, potassium salt, ammonium salt, tetraethylammonium salt, methylammonium salt, dimethylammonium salt, and ethanolamine salt.

In another preferred embodiment, the compound is deuterium substituted or isotopically labeled. Deuterium substituted compounds are capable of increasing the half-life of the compound while replicating the activity of the original compound.

In a preferred embodiment, the compound of formula 1 is a compound represented by formula 1-1, formula 1-2, formula 2-1, or formula 2-2:

wherein R is₁And R is at least one selected from deuterium, halogen, amino, hydroxyl, C1-3 alkoxy, C6-12 aryloxy, amido, substituted or unsubstituted C1-3 alkanoyloxy, C6-C12 aroyloxy, C1-10 carboxyl and substituted or unsubstituted C1-C10 alkyl respectively,

the substituent of the substituted C1-10 alkyl is selected from at least one of deuterium, halogen, hydroxyl, amino, amido, C1-3 alkanoyloxy and C1-10 carboxyl, preferably, R₁Is amino, R is at least one of hydroxyl, C1-3 alkanoyloxy and C1-10 alkyl substituted with deuterium, hydroxyl or C1-3 alkanoyloxy, the substituent of the substituted C1-3 alkanoyloxy being selected from deuterium and C6-C12 aryl,

m is an integer of 1 to 3, and n is an integer of 1 to 3.

In a preferred embodiment, the compound of formula 1 is a compound represented by formula 1-3 or formula 2-3,

wherein R is as defined above and n is an integer of 1 to 3.

In a preferred embodiment, the compound of formula 1 is selected from compound 1-1 to compound 1-6, the compound of formula 2 is selected from compound 2-6 to 2-10, preferably compound 1-2:

in another preferred embodiment, the compound is selected from the group consisting of compound 2-1 to compound 2-5 below.

In compounds 2 to 5, "AA" refers to the amino acid residue, i.e., the residue of 20 natural amino acids from which carboxyl groups have been removed.

In a preferred embodiment, wherein the viral hepatitis is hepatitis B or hepatitis D.

In a preferred embodiment, wherein the medicament is capable of reducing Hepatitis B Virus (HBV) load, HBsAg and/or HBeAg levels.

In a preferred embodiment, wherein the medicament further comprises one or more additional therapeutic or prophylactic agents, preferably, the additional therapeutic or prophylactic agent is selected from interferon, pegylated interferon, nitazoxanide or an analogue thereof, a compound of formula A or a nucleoside analogue,

preferably, the nucleoside analogue is selected from entecavir, tenofovir disoproxil fumarate and tenofovir alafenamide.

In a preferred embodiment, wherein the medicament is formulated for administration by a route selected from the group consisting of: oral, rectal, nasal, pulmonary, topical, buccal and sublingual, vaginal, parenteral, subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural, preferably oral administration, more preferably in the form of tablets or capsules.

The invention also provides a pharmaceutical composition for treating or preventing viral hepatitis, which comprises a therapeutically effective amount of the compound of the general formula 1 or a pharmaceutically acceptable salt thereof and optionally one or more additional therapeutic agents or prophylactic agents, and a pharmaceutically acceptable carrier, wherein preferably, the compound of the general formula 1 is selected from the compounds shown in formula 2, formula 2A, formula 3 or formula 4. More preferably, the additional therapeutic or prophylactic agent is selected from at least one of an interferon, a pegylated interferon, nitazoxanide or an analog thereof, a compound of formula a, or a nucleoside analog.

The invention also provides a compound for treating or preventing viral hepatitis, wherein the compound is represented by the general formula 1. Preferably the compound is compound 1-1 to compound 1-6 or compound 2-6 to compound 2-10, more preferably compound 2-1 to 2-5.

The technical scheme of the invention has the following beneficial effects:

1. the compound of the general formula 1 or the pharmaceutically acceptable salt thereof is applied to treating or preventing viral hepatitis, thereby providing a novel viral hepatitis treatment option, and the key is that the compound can remove HBsAg and HBeAg, and is expected to achieve the effect of functionally curing hepatitis B.

2. The compound of the general formula 1 or the pharmaceutically acceptable salt thereof can simultaneously and effectively reduce the load of Hepatitis B Virus (HBV), the HBsAg and/or the HBeAg level, and is expected to eliminate hepatitis B virus, cure hepatitis B and avoid the pain of lifelong medicine taking under the condition of being combined with the existing nucleoside analogue medicines.

3. As the marketed drug decitabine, the compound 1-2 or the pharmaceutically acceptable salt thereof has excellent clinical safety and pharmacokinetic properties and better drug forming property.

4. The compounds of formula 1 or pharmaceutically acceptable salts thereof can optionally be combined with one or more additional therapeutic or prophylactic agents, thereby providing a broad concept for subsequent combination drug design with potential for synergy.

Drawings

FIG. 1 shows the inhibitory effect of compounds according to the present invention on HBV DNA.

FIG. 2 is a graph showing the results of the inhibition of HBsAg by the compounds according to the examples of the present invention.

FIG. 3 shows the results of the inhibition of HBeAg by the compounds according to the examples of the present invention.

Detailed Description

In one aspect, the present invention provides the use of a compound of formula 1 or formula 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prevention of viral hepatitis,

q, X, Y and Z are each selected from C, N, O and S,

m is an integer of 1 to 2, n is an integer of 1 to 3,

wherein R is₁And R is independently selected from deuterium, halogenAt least one of amino, hydroxyl, C1-3 alkoxy, C6-12 aryloxy, amido, substituted or unsubstituted C1-3 alkanoyloxy, C6-C12 aroyloxy, C1-10 carboxyl and substituted or unsubstituted C1-C10 alkyl,

m is an integer of 1 to 3, and n is an integer of 1 to 3.

wherein R is as defined above and n is an integer of 1 to 3.

The present invention also provides a pharmaceutical composition for treating or preventing viral hepatitis comprising a therapeutically effective amount of a compound of formula 1 or formula 2 or a pharmaceutically acceptable salt thereof and optionally one or more additional therapeutic or prophylactic agents, and a pharmaceutically acceptable carrier, preferably, the compound of formula 1 is selected from the group consisting of compounds 1-1 to 1-6 and compounds 2-1 to 2-10. More preferably, the additional therapeutic or prophylactic agent is selected from at least one of an interferon, a pegylated interferon, nitazoxanide or an analog thereof, a compound of formula a, or a nucleoside analog.

In a preferred embodiment, the compound is compound 1-2. The compound 1-2 is known drug decitabine, which is an antitumor drug 5-aza-2' -deoxycytidine nucleus produced by the company Pharmachemie B.V. Netherlands and has the chemical name of 4-amino-1- (2-deoxy-beta-D-erythro-ribofuranose) -1, 3, 5-triazin-2 (1H) -one. FDA approved it for the treatment of primary and secondary myelodysplastic syndromes in 2006 in the united states. Decitabine as a specific DNA methylation transferase inhibitor can reverse the DNA methylation process, induce the tumor cells to differentiate into normal cells or induce the tumor cells to die. In tumor cells, decitabine is phosphorylated by deoxycytidine kinase and incorporated as phosphate with DNA. The high-concentration decitabine can inhibit DNA synthesis induced cell death and play a role in cytotoxicity; the incorporation of decitabine at low concentrations can replace cytosine in tumor cells to covalently bind DNA methyltransferase, inactivating DNA methyltransferase without causing cell death. At present, no report is available for treating hepatitis B.

The inventor of the application unexpectedly discovers that the series of compounds have potential activity for treating hepatitis B after analyzing and researching big data of drug structures and targets through an artificial intelligence system. After a series of biological experiments, the compound of formula 1 with the effect of treating hepatitis B is obtained.

Definition of substituents

As used herein, "alkyl" refers to a straight or branched chain aliphatic saturated hydrocarbon monovalent group having 1 to 10 carbon atoms, and non-limiting examples thereof include methyl, ethyl, propyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, heptyl, octyl, nonyl, decyl, and the like. As used herein, "hydroxy" refers to an-OH group that may be protected by a protecting group, for example, by an ester linkage to form an alkanoyloxy, e.g., acetoxy, protecting group. Under the action of in vivo esterase, the protecting group can be removed to form active hydroxyl. As used herein, "halogen" refers to at least one of fluorine, chlorine, bromine, iodine. As used herein, "amino" refers to substituted or unsubstituted-NH₂Groups, which can be protected by amide bond formation and act by in vivo enzymatic degradation during metabolism to form active amino groups. As used herein, "amide group" refers to a group having an amide bond formed after protecting an amino group. For example, an amide bond formed by dehydration reaction of a carboxyl group of an amino acid such as alanine with an amino group.

As used herein, "alkanoyloxy" refers to the group RCOO-such as acetoxy, propionyloxy, butyryloxy, etc., formed after the hydroxyl group is protected by an ester bond, wherein R may be C1-10 alkyl.

As used herein, "carboxy" refers to-RCOOH, and R may be a C1-10 alkyl group.

Viral hepatitis

The etiological typing of viral hepatitis is currently recognized by five hepatitis viruses, namely hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus and hepatitis E virus, which are respectively written as HAV, HBV, HCV, HDV and HEV, and the rest are RNA viruses except the hepatitis B virus which is a DNA virus.

Hepatitis b is an infectious disease mainly caused by hepatitis b virus, and is a liver disease. Clinically, the symptoms of anorexia, nausea, epigastric discomfort, liver pain and hypodynamia are mainly manifested. Some patients may have jaundice fever and hepatomegaly with impaired liver function. Some patients can become chronic, even develop cirrhosis of the liver, and a few can develop liver cancer.

The etiological agent of hepatitis b is hepatitis b virus, abbreviated as HBV, which is DNA virus. The genome is a double-stranded, circular, incompletely closed DNA. The outermost layer of the virus is the outer membrane or coat membrane of the virus, the inner layer is the core part, and the nucleoprotein is the core antigen (HBcAg) and cannot be detected in the serum. Serum from HBsAg positive patients was observed under electron microscope to show 3 kinds of particles, circular and filamentous particles with a diameter of 22nm, and less spherical particles with a diameter of 42 angstroms, also called Dane's particles, as complete HBV particles.

The markers for hepatitis b were detected as follows: (ii) HBsAg and anti-HBs: HBsAg positive indicates that HBV is currently in the stage of infection, and anti-HBs positive for immunoprotective antibodies indicates that immunity to HBV has developed. The diagnosis basis of the chronic HBsAg carrier is that the chronic HBsAg carrier has no clinical symptoms and physical signs, the liver function is normal, and the HBsAg is continuously positive for more than 6 months. (vii) HBeAg and anti-HBe: HBeAg positive is an index of HBV active replication and strong infectivity, and the change of the detected serum from HBeAg positive to anti-HBe positive indicates that the disease is relieved and the infectivity is weakened. ③ HBcAg and anti-HBc: HBcAg positive suggests that there is a direct reaction of complete HBV particles, and active replication of HBV is less clinically useful due to the complex detection method. anti-HBc is a marker of HBV infection, and anti-HBc IgM positive indicates that the virus is replicated in vivo at an early stage of infection. HBsAg, HBeAg and anti-HBc are all positive in chronic mild hepatitis B and HBsAg carriers, and have high infectivity index and are difficult to turn negative.

In a preferred embodiment, the medicament further comprises one or more additional therapeutic or prophylactic agents. In a preferred embodiment, the additional therapeutic or prophylactic agent is selected from an interferon or a nucleoside analogue. In a preferred embodiment, the nucleoside analogue is selected from entecavir, tenofovir disoproxil fumarate and tenofovir alafenamide.

Additional therapeutic or prophylactic agents

In some embodiments, the additional therapeutic or prophylactic agent is selected from one or more of entecavir, tenofovir disoproxil fumarate, and tenofovir alafenamide, for example, one selected from entecavir, tenofovir disoproxil fumarate, and tenofovir alafenamide or at least two selected from entecavir, tenofovir disoproxil fumarate, and tenofovir alafenamide.

Entecavir (Entecavir) is chemically known as 2-amino-1, 9-dihydro-9- [ (1S,3R,4S) -4-hydroxy-3- (hydroxymethyl) -2-methylenecyclopentane ] -6H-purin-6-one and has the following structural formula:

US patent US5206244 discloses entecavir and its use for the treatment of hepatitis b virus; a novel synthesis of entecavir is disclosed in WO 9809964; WO0164421 discloses low dose entecavir solid formulations.

Entecavir is a highly effective antiviral agent, developed by schrobo corporation in the 90 s of the 20 th century, and has a strong anti-HBV effect. It can be phosphorylated to active triphosphate, which has a half-life in cells of 15 h. Entecavir triphosphate inhibits all three activities of the viral polymerase (reverse transcriptase) by competing with deoxyguanosine triphosphate, the natural substrate of HBV polymerase: (1) the start of HBV polymerase; (2) formation of a reverse transcribed negative strand of a pregenomic mRNA; (3) synthesis of HBV DNA plus strand.

Tenofovir disoproxil fumarate (the name of England: (TDF); (R) - [ [2- (6-amino-9H-purin-9-yl) -1-methylethoxy ] methyl ] phosphonic acid diisopropoxycarbonylmethyl ester fumarate) is an ester precursor of Tenofovir, belongs to a novel nucleotide reverse transcriptase inhibitor, and has the activity of inhibiting HBV viruses.

TDF is another novel open-ring nucleoside phosphonate successfully developed by Gilidard company in the United states following Adefovir dipivoxil, is first marketed in the United states in 10 months in 2001, and is currently marketed in countries such as Europe, Australia, and Canada.

TDF inhibits viral polymerase in vivo by competitively binding to the natural deoxyribose substrate and terminates DNA strand synthesis by insertion into DNA. The main action mechanism is that the tenofovir is hydrolyzed into tenofovir after being orally taken, the tenofovir is phosphorylated by cell kinase to generate a metabolite tenofovir diphosphate with pharmacological activity, the tenofovir diphosphate competes with 5 '-triphosphate deoxyadenosine monophosphate to participate in the synthesis of virus DNA, and after entering the virus DNA, the DNA is prevented from being prolonged due to the lack of 3' -OH groups, so that the replication of the virus is blocked. Clinical application shows that TDF has obvious curative effect on HBV virus and less toxic side effect, so that TDF has wide clinical application foreground.

Tenofovir Alafenamide (Tenofovir Alafenamide), a prodrug of the new Nucleoside Reverse Transcriptase Inhibitor (NRTI) Tenofovir (Tenofovir) developed by Gilidard scientific, USA. Compared with the prior generation of similar anti-hepatitis B medicine tenofovir disoproxil TDF, the antiviral activity of tenofovir alafenamide is 10 times, the stability in blood plasma is 200 times, and the half-life period is improved by 225 times. Compared with TDF, the tenofovir alafenamide only needs one tenth of TDF administration dosage to achieve the same antiviral curative effect as TDF. Therefore, the tenofovir alafenamide is used for preventing or/and treating Hepatitis B Virus (HBV) infection and has better curative effect, higher safety and lower drug resistance.

In addition to the above active agents, the medicaments or pharmaceutical compositions described herein may optionally comprise one or more additional other agents useful in the treatment of HBV, such as, but not limited to, 3-dioxygenase (IDO) inhibitors, antisense oligonucleotides targeted to viral mRNA, apolipoprotein a1 modulators, arginase inhibitors, B-and T-lymphocyte attenuating agent inhibitors, Bruton's Tyrosine Kinase (BTK) inhibitors, CCR2 chemokine antagonists, CD137 inhibitors, CD160 inhibitors, CD305 inhibitors, CD4 agonists and modulators, compounds targeted to HBcAg, compounds targeted to hepatitis B core antigen (HBcAg), covalently closed circular DNA (cccdna) inhibitors, cyclophilin inhibitors, cytokines, cytotoxic T-lymphocyte-associated protein 4(ipi4) inhibitors, DNA polymerase inhibitors, endonuclease modulators, epigenetic modifiers, farnesoid X receptor agonists, gene modifiers or editors, HBsAg inhibitors, HBsAg secretion or assembly inhibitors, HBV antibodies, HBV DNA polymerase inhibitors, HBV replication inhibitors, HBV rnase inhibitors, HBV vaccines, HBV viral entry inhibitors, HBx inhibitors, hepatitis b large envelope protein modulators, hepatitis b large envelope protein stimulators, hepatitis b structural protein modulators, hepatitis b surface antigen (HBsAg) inhibitors, hepatitis b surface antigen (HBsAg) secretion or assembly inhibitors, hepatitis b virus E antigen inhibitors, hepatitis b virus replication inhibitors, hepatitis virus structural protein inhibitors, HIV-1 reverse transcriptase inhibitors, hyaluronidase inhibitors, IAP inhibitors, IL-2 agonists, IL-7 agonists, immunoglobulin G modulators, immunomodulators, indoleamine-2, ribonucleotide reductase inhibitors, interferon agonists, interferon alpha 1 ligands, interferon alpha 2 ligands, interferon alpha 5 ligand modulators, interferon alpha ligands, interferon alpha ligand modulators, interferon alpha receptor ligands, interferon beta ligands, interferon receptor modulators, interleukin-2 ligands, ipi4 inhibitors, lysine demethylase inhibitors, histone demethylase inhibitors, KDM5 inhibitors, KDM1 inhibitors, a member of the lectin-like receptor subfamily G1 inhibitor, lymphocyte activation gene 3 inhibitors, lymphotoxin beta receptor activators, microRNA (miRNA) gene therapeutics, Axl modulators, B7-H3 modulators, B7-H4 modulators, CD160 modulators, CD161 modulators, CD27 modulators, CD47 modulators, CD70 modulators, GITR modulators, HEVEM modulators, ICOS modulators, Mer modulators, NKG2A modulators, NKG2D modulators, OX40 modulators, SIRPa modulators, TIGIT modulators, Tim-4 modulators, Tyro modulators, Na + -taurate cotransporter (NTCP) inhibitors, natural killer cell receptor 2B4 inhibitors, NOD2 gene stimulators, nucleoprotein inhibitors, nucleoprotein modulators, PD-1 inhibitors, PD-L1 inhibitors, PEG-interferon lambda, peptidyl prolyl isomerase inhibitors, phosphatidylinositol-3 kinase (PI3K) inhibitors, recombinant Scavenger Receptor A (SRA) proteins, recombinant thymosin alpha-1, retinoic acid inducible gene 1 stimulators, reverse transcriptase inhibitors, ribonuclease inhibitors, RNA polymerase inhibitors, short interfering RNA (siRNA), short synthetic hairpin RNA (sshRNA), (sshRNA) inhibitors, HEVEM modulators, ICOS modulators, SIRP modulators, TIGIT modulators, TIG-4 modulators, Tyro modulators, TyCP modulators, Na + -NTCP inhibitors, natural killer-co-transporters, natural killer cells receptor 2B4 inhibitors, NOD2 gene stimulators, nuclear protein inhibitors, PD-1 inhibitors, PD-induced gene 1 inhibitors, short interfering RNA polymerase inhibitors, short synthetic hairpin RNA (siRNA) and the like, SLC10a1 gene inhibitors, SMAC mimetics, Src tyrosine kinase inhibitors, interferon gene Stimulator (STING) agonists, NOD1 stimulators, T cell surface glycoprotein CD28 inhibitors, T cell surface glycoprotein CD8 modulators, thymosin agonists, thymosin alpha 1 ligands, Tim-3 inhibitors, TLR-3 agonists, TLR-7 agonists, TLR-9 agonists, TLR9 gene stimulators, toll-like receptor (TLR) modulators, viral ribonucleotide reductase inhibitors, zinc finger nucleases or synthetic nucleases (TALENs) and combinations thereof.

As used herein, "therapeutically effective amount" or "effective amount" refers to an amount that is effective at a dose and for a period of time required to achieve a desired therapeutic result. A therapeutically effective amount of a therapeutic agent for hepatitis b will depend on the nature of the disorder or condition and on the particular agent, and can be determined by standard clinical techniques known to those skilled in the art.

The therapeutic result can be, for example, alleviation of symptoms, prolongation of survival, improvement of quality of life, and the like. The therapeutic result need not be a "cure". The therapeutic outcome may also be prophylactic. The most preferred therapeutic effects are functional healing and clearance of hepatitis b virus.

In a preferred embodiment, the medicament is formulated for administration by a route selected from the group consisting of: oral, rectal, nasal, pulmonary, topical, buccal and sublingual, vaginal, parenteral, subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural.

In a preferred embodiment, the medicament is formulated for oral administration, preferably in the form of a tablet or capsule.

Route of administration

The medicaments or pharmaceutical compositions of the present disclosure are administered by any route suitable for the condition to be treated. Suitable routes include oral, rectal, nasal, pulmonary, topical (including buccal and sublingual), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) and the like. In certain embodiments, the medicament or pharmaceutical composition disclosed herein is administered by intravenous injection. It will be appreciated that the preferred route may vary depending on, for example, the condition of the recipient. One advantage of the disclosed medicaments or pharmaceutical compositions is that they are orally bioavailable and can be administered orally.

Pharmaceutical composition

In certain embodiments, the compound of formula 1 or the compound drug of formula 2, or a pharmaceutically acceptable salt thereof, is administered in a pharmaceutical composition. The pharmaceutical compositions of the present disclosure may be formulated with conventional carriers and excipients, which will be selected in accordance with common practice. Tablets will contain excipients, glidants, fillers, binders and the like. Aqueous formulations are prepared in sterile form and, when used for delivery by non-oral administration, are generally isotonic. All formulations will optionally contain Excipients such as those described in the Handbook of Pharmaceutical Excipients (1986). Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkyl cellulose, hydroxyalkyl methyl cellulose, stearic acid, and the like. The pH of the formulation ranges from about 3 to about 11, but is typically from about 7 to 10. In some embodiments, the pH of the formulation ranges from about 2 to about 5, but typically from about 3 to 4.

The formulations include those suitable for the aforementioned routes of administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Techniques and formulations are commonly found in Remington's Pharmaceutical Sciences (Mack Publishing co., Easton, PA). Such methods include the step of bringing into association the active ingredient with the carrier which is composed of one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then shaping the product as necessary.

Formulations of the invention suitable for oral administration may exist as follows: discrete units, such as capsules or tablets, each containing a predetermined amount of active ingredient; a powder or granules; solutions or suspensions in aqueous or non-aqueous liquids; or an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.

Tablets are made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by: the active ingredient in a free-flowing form such as a powder or granules is compressed in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersant. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. The tablets may optionally be coated or scored and optionally formulated so as to provide sustained or controlled release of the active ingredient therefrom.

Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive oil.

The pharmaceutical compositions of the present disclosure may also be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. The suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol, or as a lyophilized powder. Acceptable carriers and solvents that may be employed include water, ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Acceptable carriers and solvents that may be employed include water, ringer's solution, isotonic sodium chloride solution and hypertonic sodium chloride solution.

Additional objects, advantages and novel features of the present invention will become apparent to one of ordinary skill in the art upon examination of the following examples.

Examples

Example 1 evaluation of in vitro anti-HBV Activity of Compounds 1-2 (Decitabine) Using HepG2-NTCP cells

Materials and methods

Cell: HepG2-NTCP cells were provided by Shanghai medicine Mingkude New drug development Co., Ltd.

Compounds 1-2: purchased from Shanghai ceramic Biochemical technology, Inc.

The control compounds RG7834 and ETV (20mM DMSO solution) were provided by Shanghai drug Mingkuda New drug development, Inc. The compound information is shown in table 1.

TABLE 1 Compound information Table

Virus: HBV type D was concentrated from HepDES19 cell culture supernatant and is detailed in table 2.

TABLE 2 Virus information Table

GE: genome equivalents

Reagent

The main reagents include QIAamp96 DNA kit (cat # Qiagen-51162), CellTiter-Glo (cat # G7558), HBsAg ELISA kit (cat # Antu-CL0310), HBeAg ELISA kit (cat # Antu-CL 0312) and FastStart Universal Probe Master (cat # Roche-04914058001).

Instrument for measuring the position of a moving object

The main instruments used in this study were a fluorescent quantitative PCR instrument (7900HT Fast Real-Time PCR System/QuantStaudio (TM)6Flex System) and a microplate reader (BioTek Synergy 2).

The compound preparation method comprises the following steps:

for the preparation of 20mM concentration, the volume of the solvent DMSO (. mu.l) is the sample mass (mg). times.purity ÷ molecular weight ÷ 20X 10⁶

Control compounds include ETV (batch No.: P1214012; 99.0% purity) available from Shanghai Tantake Technology, Inc. The positive control compound RG7834 (batch No.: ET 25747-14-P1; 99.5% purity) was purchased from Shanghai drug Mingkuda, New drug development, Inc. The mother liquors of the above control compounds were all at 20mM concentration and stored at-20 ℃.

TABLE 3 Main Agents and cellular Virus List

Experimental protocol

Plating cells and compound treatment

On day 0, HepG2-NTCP was plated into 48-well plates (7.5X 10)⁴Individual cells/well).

On day 2, cells were pretreated with the compound for 2 hours before the addition of D-type HBV to infect HepG2-NTCP cells (compound was added at the same time as infection). Test compounds were diluted to 3 single drug concentrations, 1 combination drug concentration (compound of formula 2 + ETV), 2 duplicate wells. The control compound was ETV. The concentrations of the three single drugs were 1, 10 and 20. mu.M, respectively, and the concentration of the combination was 20. mu.M + ETV 0.1nM for the test compound. Control compounds were at 7 concentrations of ETV and RG7834 each, with wells containing only DMSO and no compound set up and ETV single drug concentration 0.1nM, 2 duplicate wells tested.

Fresh medium containing compounds was replaced on days 3, 5 and 7.

On day 9, supernatants were collected and the collected cell supernatants were assayed for HBeAg and HBsAg by ELISA and HBV DNA levels by qPCR. Meanwhile, CellTiter-Glo detects cell viability, and the collected cells are frozen and stored (for later use).

Sample detection

1) qPCR method for detecting HBV DNA content in cell culture supernatant

DNA was extracted from the cell culture supernatant according to the QIAamp96 DNA Blood Kit instructions. And detecting the content of HBV DNA by the HBV specific primer qPCR. And (3) PCR reaction: at 95 ℃ for 10 min; 95 ℃, 15sec, 60 ℃, 1min, 40 cycles.

2) ELISA method for detecting content of HBeAg and HBsAg in cell culture supernatant

The method refers to the kit specification, and the method is briefly described as follows: respectively taking 50 mu l of standard substance, sample and reference substance, adding 50 mu l of enzyme conjugate into each hole, incubating for 60 minutes at 37 ℃, washing the plate with washing liquor, sucking dry, adding 50 mu l of premixed luminescent substrate, incubating for 10 minutes at room temperature in a dark place, and finally measuring the luminescent value by an enzyme-linked immunosorbent assay.

3) CellTiter-Glo cell viability assay

Cell viability was determined with reference to CellTiter-Glo kit instructions, the method is briefly as follows: after collection of the cell culture supernatant, CellTiter-Glo (medium 1:1 dilution) was added to each well, incubated at room temperature for 10 minutes, and the luminescence was measured by a microplate reader (results not shown).

Data analysis

HBV DNA inhibition (%) was (1-HBV copy number of sample of compound group/HBV copy number of DMSO group) × 100%

The Hbe/sAg inhibition rate (%) was (1-Hbe/sAg value of sample/DMSO control Hbe/sAg value) × 100%

Analysis of results

The results of the measurements are shown in tables 4-6 and FIGS. 1-3.

TABLE 4 HBV DNA inhibition by test Compounds

TABLE 5 HBsAg inhibition of test Compounds

TABLE 6 HBeAg inhibition of the test compounds

The above experimental results show that, compared with the blank control group, the compound groups 1-2 can effectively reduce the HBV viral load, and can simultaneously reduce HBeAg and HBsAg by 34.74 and 50.78% respectively under the condition of reducing HBV DNA by 80.51%. While entecavir can only reduce HBV DNA as reported in the literature, it has no substantial effect on reducing HBeAg and HBsAg. As can be seen, the compound 1-2 can effectively reduce HBeAg and HBsAg compared with entecavir, thereby hopefully eliminating hepatitis B virus and achieving functional cure.

Especially the inhibiting effect on HBsAg and HBeAg, so that the compound can be used as a candidate drug for functionally curing hepatitis B and eliminating hepatitis B virus, especially when the compound is combined with nucleoside analogue drugs, such as entecavir and TAF, which can reduce the HBV titer but can not reduce the HBsAg and HBeAg, for example, when the nucleoside analogue drugs are used for reducing the virus titer, so that the virus content in human bodies is reduced to a lower level, and decitabine is used for further reducing the HBsAg and HBeAg, the virus can be further eliminated, and even the complete cure can be achieved.

While the invention has been described with reference to specific embodiments, those skilled in the art will recognize that changes or modifications can be made to the described embodiments without departing from the spirit and scope of the invention, which is defined by the appended claims.

Claims

1. Use of a compound of formula 1 or formula 2 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or prevention of viral hepatitis,

q, X, Y and Z are each selected from C, N, O and S,

m is an integer of 1 to 2, n is an integer of 1 to 3,

2. The use according to claim 1, wherein the compound of formula 1 or formula 2 is a compound represented by formula 1-1, formula 1-2, formula 2-1, or formula 2-2, respectively:

formula 1-1

Formula 1-2

Formula 2-1

Formula 2-2

m is an integer of 1 to 2, and n is an integer of 1 to 3.

3. The use according to claim 1, wherein the compound of formula 1 or formula 2 is a compound represented by formula 1-3 or formula 2-3, respectively,

formulas 1 to 3

Formula 2-3

Wherein R is as defined in claim 1, and n is an integer of 1 to 3.

4. The use according to claim 1, wherein the compound of formula 1 is selected from the group consisting of compounds 1-1 to 1-6, the compound of formula 2 is selected from the group consisting of compounds 2-6 to 2-10, preferably compounds 1-2:

5. the use of any one of claims 1-4, wherein the viral hepatitis is hepatitis B or hepatitis D.

6. The use of claim 5, wherein the medicament is for reducing Hepatitis B Virus (HBV) load, HBsAg and/or HBeAg levels.

7. The use of any one of claims 1-4, wherein the medicament further comprises one or more additional therapeutic or prophylactic agents, preferably the additional therapeutic or prophylactic agent is selected from at least one of an interferon, a PEGylated interferon, nitazoxanide or an analogue thereof, a compound of formula A, or a nucleoside analogue,

formula A

8. The use of any one of claims 1-7, wherein the medicament is formulated for administration by a route selected from the group consisting of: oral, rectal, nasal, pulmonary, topical, buccal and sublingual, vaginal, parenteral, subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural, preferably oral administration, more preferably in the form of tablets or capsules.

9. A pharmaceutical composition for the treatment or prevention of viral hepatitis comprising a therapeutically effective amount of a compound of formula 1 or a pharmaceutically acceptable salt thereof and optionally one or more additional therapeutic or prophylactic agents, preferably the compound of formula 1 is selected from a compound of formula 2A or formula 2, more preferably the additional therapeutic or prophylactic agent is selected from at least one of an interferon, a pegylated interferon, nitazoxanide or an analogue thereof, a compound of formula a or a nucleoside analogue, and a pharmaceutically acceptable carrier.

10. A compound for treating or preventing viral hepatitis, wherein the compound is represented by formula 1, preferably a compound represented by formula 2, formula 2A, formula 3 or formula 4, more preferably a deuterium-substituted compound represented by formula 2, formula 2A, formula 3 or formula 4.