Hepatitis B virus receptor OATP and application thereof

Abstract

The invention provides a hepatitis B virus receptor OATP and application thereof. Specifically, the invention provides an application of a substance in preparation of a medicament for preventing and / or treating animal hepatitis B virus (HBV) and / or hepatitis D virus (HDV) infection or diseases related to the infection, the substance prevents or reduces the interaction between HBV and / or HDV and an organic anion transporting polypeptide (OATP) in the animal and / or prevents or reduces the expression and / or function of OATP in the animal. The invention also provides an HBV and / or HDV susceptible cell model and animal model obtained by transferring an exogenous SLCO gene or expressing an exogenous OATP protein, an establishment method of the cell model and animal model, a drug delivery method and carrier for targeting OATP, and an OATP targeting delivery polypeptide carrier derived from a hepatophilic DNA virus surface antigen.

Description

Hepatitis B virus receptor OATP and its application Technical Field

[0001] The present invention relates to HBV receptor OATPs and their applications, specifically to their use in the treatment and prevention of HBV and / or HDV infection and related diseases, in HBV and / or HDV cell and animal infection models, and in drug screening. The present invention also relates to the use of polypeptides derived from hepadnavirus surface antigens in the treatment and prevention of HBV and / or HDV infection and related diseases, in drug delivery vehicles targeting OATPs, and in inhibiting OATP transport function. Background Art

[0002] Approximately 350 million people worldwide have been infected with hepatitis B virus (HBV), and approximately 200 million have chronic HBV. These individuals are at increased risk for developing cirrhosis, liver failure, and liver cancer. HBV belongs to the Hepadnaviridae family, an ancient pathogen that naturally infects mammals (Orthohepadnavirus, OHV), birds (Avihepadnavirus, AHV), fish (Metahepadnavirus), and amphibians (Herpetohepadnavirus). Among mammals, rodent Orthohepadnaviruses have been found only in three species of the Sciuridae family: the arctic ground squirrel, the ground shrew, and the woodchuck. In primates, protohepadnaviruses have been isolated from hominoid species, including humans, chimpanzees, gorillas, gibbons, and orangutans. Natural protohepadnavirus infections have been reported in three New World monkey species, including woolly monkeys, capuchin monkeys, and squirrel monkeys. Natural infection with bat protohepadnaviruses infects multiple species in at least five clades [Jacquet S, et al. J Virol. 2019; 93(5): e01738-18.].

[0003] The human HBV genome consists of partially double-stranded circular DNA, approximately 3.2 kb in length. Four highly overlapping gene regions encode at least eight proteins: the S region encodes the large (L), medium (M), and small (S) surface antigens HBs; the C gene encodes pre-C, HBc, and HBsinus; the P gene encodes the polymerase; and the X gene encodes HBx. HBV-related particles primarily have three morphologies: Dane particles with a diameter of 42-47 nm; 20 nm spherical particles that are 10,000-1,000,000 times more abundant than Dane particles; and a small number of filamentous fibers with a diameter of 20 nm and varying lengths. Dane particles are the only infectious viral particles of HBV, composed of genomic DNA, viral DNA polymerase, nucleocapsid, and envelope. The HBV viral envelope contains three surface antigenic proteins: large (L), medium (M), and small (S). These proteins are encoded by a single open reading frame in the S region gene with three distinct translation start sites: L (Pre-S1+Pre-S2+S), M (Pre-S2+S), and S (S). The expression levels of these three surface proteins vary significantly, and their distribution patterns differ significantly. S protein is highly expressed and is the predominant HBs protein in the circulating blood of HBV-infected patients, while M and L proteins are present in very low concentrations, accounting for only 5–15% and 1–2% of the total HBs protein, respectively. The envelope proteins of 20 nm spherical particles and filamentous fibers are primarily composed of S protein and varying amounts of M protein, with virtually no L protein present. In contrast, the envelope proteins of Dane particles are primarily L protein.

[0004] Hepatitis D virus (HDV) is a defective RNA virus and a satellite virus of HBV, present as spherical particles approximately 36 nm in diameter. Its interior consists of a nucleocapsid composed of a 1.7 kb circular RNA and approximately 200 copies of the hepatitis D antigen (HDAg); its outer envelope contains the HBV surface antigen and the host cell plasma membrane. HDV can only be propagated in the presence of HBV; it rarely infects the host cell alone. Co-infection with HBV and HDV often leads to severe clinical outcomes. An estimated 15 million people worldwide are chronically infected with HDV, with rates of cirrhosis and liver cancer three times and nine times higher than those of chronic hepatitis B, respectively. It is considered the most severe form of viral hepatitis. Since HDV uses the HBV envelope, the infection receptors of HBV and HDV are considered to be very similar [Hughes et al., Lancet 378, 73 (2011); Sureau, Curr Top Microbiol Immunol 307, 113 (2006); Barrera et al., J Virol 78, 5233 (2004)].

[0005] The Pre-S1 region of the HBV surface antigen contains a key amino acid sequence that binds to hepatocyte-specific receptors [Le Seyec J et al. J Virol. 1999; 73(3): 2052-7.], among which the N-terminal amino acid sequence and myristic acid modification are essential for HBV infection [Bruss V et al. Virology. 1996; 218: 396-399]. The human hepatocyte cell line HepG2 can support HBV and HDV infection after being transfected with human sodium taurocholate cotransporting polypeptide (hNTCP), and the HBVPreS1 region polypeptide with N-terminal myristic acid modification can bind to hNTCP. Therefore, hNTCP is considered to be a functional receptor for HBV and HDV infection [Yan H et al. Elife. 2012; 1: e00049.; Ni Y et al. Gastroenterology. 2014; 146(4): 1070-83.]. However, HepG2 cells transfected with hNTCP (HepG2-hNTCP) can only be infected with HBV in the presence of polyethylene glycol (PEG) [Yan H et al. Elife. 2012; 1: e00049.], and the infection is not a natural process and has a low infection efficiency [Herrscher C et al. Cells. 2020 Jun 18; 9(6): 1486]. Moreover, non-hepatic human cell lines or non-human hepatocytes (such as rat, mouse, and dog hepatocytes) do not support HBV infection even if they are transfected with hNTCP [Lempp FA et al. Hepatology. 2017; 66(3): 703-716]; hNTCP transgenic mice also do not support HBV infection [He W et al. J Virol. 2016 Sep 12; 90(19): 8866-74]. It can be seen that hNTCP is a necessary but not sufficient condition for efficient HBV infection, and other components are involved in the formation of the receptor [Herrscher C et al. Cells. 2020 Jun 18; 9(6): 1486]. Except for humans, tree shrews, chimpanzees and their primary hepatocytes that are naturally susceptible to HBV or HDV, cell and animal models that can be naturally infected with HBV are currently unavailable. Therefore, it is necessary to further study other components of the HBV receptor and develop effective methods for the treatment and prevention of HBV and / or HDV infection and related diseases. It is also necessary to develop in vitro cell infection systems and transgenic animal models for screening drugs that can treat and prevent HBV and / or HDV. The mechanism of HBV infection of hepatocytes can also be used to develop drug delivery vectors targeting the liver.

[0006] Organic anion transporting polypeptides (OATPs) are encoded by the Solute Carrier Organic Anion Transporter Family Member (SLCO) gene. Over 300 OATP (SLCO) transporters have been identified in over 40 species, with at least 11 members of this superfamily expressed in humans. OATPs are typically divided into families and subfamilies based on amino acid sequence homology of 40% and 60%. Six families (OATP1-6) and 13 subfamilies (e.g., OATP1A, OATP1B, OATP1C, etc.) are identified in humans, monkeys, dogs, and rodents (Hagenbuch B, et al. Mol Aspects Med. 2013 Apr-Jun;34(2-3):396-412). OATP is predicted to be a 12-transmembrane protein, a sodium-independent transporter with a wide range of endogenous and exogenous transport substrates, mainly amphipathic organic anions.

[0007] Summary of the Invention

[0008] The present invention provides a method and agent for preventing and / or treating hepatitis B virus (HBV) and / or hepatitis D virus (HDV) infection, or diseases associated with such infection, in animals. The method comprises administering to an animal in need of such prevention and / or treatment an effective amount of an agent that can prevent or reduce the interaction between HBV and / or HDV and an organic anion transporting polypeptide (OATP) in the animal, and / or can prevent or reduce the expression and / or function of the OATP in the animal.

[0009] In one or more embodiments, the animal is preferably a mammal or a bird, more preferably a human, a chimpanzee, a bonobo, a tree shrew, a duck, a marmot, an arctic shrew, a shrew, a bat, a woolly monkey, a capuchin monkey, and a squirrel monkey. More preferably, the animal is a human.

[0010] In one or more embodiments, the OATP includes but is not limited to mammalian, avian, fish, and amphibian OATPs, preferably mammalian and avian OATPs, more preferably mammalian and avian OATP1B subfamily, including OATP1B3 and / or OATP1B1 and / or OATP1B2, more preferably mammalian OATP1B3 and / or OATP1B1 and / or OATP1B2, and even more preferably OATP1B3 and / or OATP1B1 and / or OATP1B2 from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchins, and squirrel monkeys. Even more preferably, the OATP is human OATP1B3 and / or human OATP1B1.

[0011] In one or more embodiments, the agent that prevents or reduces the interaction between HBV and / or HDV and OATP can act on HBV and / or HDV, or on OATP, or on both HBV and / or HDV and OATP; preferably, the agent acts on HBV and / or HDV, or on OATP; further preferably, the agent acts on human OATP; even more preferably, the agent acts on human OATP1B3 and / or human OATP1B1, that is, it targets human OATP1B3 and / or human OATP1B1.

[0012] In one or more embodiments, the agent that prevents or reduces the interaction between HBV and / or HDV and OATP is preferably a polypeptide, antibody, protein, small molecule agent, polynucleotide, or other agent that can interact with OATP. Further preferably, the agent is a small molecule agent, protein, antibody, or polypeptide that can interact with human OATP1B3 and / or human OATP1B1, and even more preferably, an antibody or polypeptide that can interact with human OATP1B3 and / or human OATP1B1, and even more preferably, a polypeptide that can interact with human OATP1B3 and / or human OATP1B1. Even more preferably, the agent is a polypeptide derived from the amino acid sequence of the HBVPre-S1 region, with a fatty acid modification at its N-terminus. Even more preferably, the agent is a polypeptide derived from the amino acid sequence of positions 13-59 of the human genotype C HBVPre-S1 region, with a fatty acid modification at its N-terminus. More preferably, the agent is a polypeptide represented by SEQ ID NO: 67 and SEQ ID NO: 68, which are respectively modified with myristic acid at the N-terminus and amidated at the C-terminus.

[0013] In one or more embodiments, the agent that can prevent or reduce the expression and / or function of OATP in the animal is preferably an agent that can act on the SLCO gene, OATP mRNA, OATP protein translation, expression, modification, transport or degradation. Further preferably, the agent is an agent that acts on gene knockout, gene insertion, homologous recombination, gene editing and transcriptional inhibition of the SLCO gene, or an antisense nucleic acid or RNA interference that acts on OATP mRNA, or a ubiquitination or protein degradation targeting chimera (PROTAC) that acts on OATP protein. Further preferably, the agent is a gene editing agent targeting the SLCO gene, an antisense nucleic acid or RNA interference that targets OATP mRNA, or a PROTAC agent that targets OATP protein. Still further preferably, the agent is a polynucleotide and an oligonucleotide, further preferably a DNA or RNA sequence that can achieve gene editing, a DNA or RNA sequence that can achieve RNA interference, a DNA or RNA sequence that can achieve antisense nucleic acid, and more preferably a small nucleotide (Small interfering RNA, siRNA) that can achieve RNA interference.

[0014] In one or more embodiments, the methods and agents for treating and / or preventing HBV and / or HDV infection or diseases associated with the infection can be used alone or in combination with agents that can prevent or reduce the interaction between HBV and / or HDV and NTCP and / or agents that can prevent or reduce the expression and / or function of NTCP.

[0015] The present invention also provides cell and animal models susceptible to HBV and / or HDV, and methods for establishing such models. The exogenous SLCO gene is transfected into cells or animals, and / or an exogenous OATP protein is expressed in cells or animals. This allows cells or animals that were previously non-susceptible or non-infected with HBV and / or HDV to become susceptible to HBV and / or HDV after SLCO gene transfection or exogenous OATP protein expression. The present invention also encompasses drugs and lead compounds screened using these HBV and / or HDV-infected cell and / or animal models. These drugs can be used to treat and / or prevent HBV and / or HDV infection.

[0016] In one or more embodiments, the SLCO gene includes, but is not limited to, SLCO genes from mammals, birds, fish, and amphibians, preferably mammalian and avian SLCO genes, more preferably mammalian and avian SLCO1B subfamily, including SLCO1B3 and / or SLCO1B1 and / or SLCO1B2. More preferably, the SLCO gene is mammalian SLCO1B3 and / or SLCO1B1 and / or SLCO1B2, and even more preferably SLCO1B3 and / or SLCO1B1 and / or SLCO1B2 from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchin monkeys, or squirrel monkeys. Even more preferably, the SLCO gene is human (e.g., as shown in SEQ ID NO: 3), chimpanzee, or tree shrew SLCO1B3 and / or human, chimpanzee, or tree shrew SLCO1B1. Still more preferably, the SLCO gene is human SLCO1B3 and / or human SLCO1B1 (eg, as shown in SEQ ID NO: 3, 6).

[0017] In one or more embodiments, the OATP protein includes but is not limited to mammalian, avian, fish, and amphibian OATP proteins, preferably mammalian and avian OATP proteins, further preferably mammalian and avian OATP1B subfamily, including OATP1B3 and / or OATP1B1 and / or OATP1B2. More preferably, the OATP protein is mammalian OATP1B3 and / or OATP1B1 and / or OATP1B2, further preferably OATP1B3 and / or OATP1B1 and / or OATP1B2 derived from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchin monkeys, or squirrel monkeys. Further preferably, the OATP protein is human, chimpanzee, or tree shrew OATP1B3, and / or human, chimpanzee, or tree shrew OATP1B1. Even more preferably, the OATP protein is human OATP1B3 and / or human OATP1B1 (e.g., as shown in SEQ ID NOs: 1 and 4).

[0018] In one or more embodiments, in the cell or animal model susceptible to HBV and / or HDV and the method for establishing the same, the SLCO gene can be transfected alone or simultaneously with an exogenous SLC10A1 gene; or an exogenous OATP protein can be expressed alone or simultaneously with an exogenous NTCP protein. The SLC10A1 gene includes, but is not limited to, mammalian, avian, fish, and amphibian SLC10A1 genes, preferably mammalian and avian SLC10A1 genes, and more preferably, SLC10A1 genes from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchins, squirrel monkeys, and ducks. More preferably, the SLC10A1 gene is a human SLC10A1 gene (e.g., as shown in SEQ ID NO: 79), a chimpanzee SLC10A1 gene, or a tree shrew SLC10A1 gene. Even more preferably, the SLC10A1 gene is a human SLC10A1 gene (e.g., as shown in SEQ ID NO: 79). The NTCP protein includes, but is not limited to, mammalian, avian, fish, and amphibian NTCP proteins, preferably mammalian and avian NTCP proteins, and more preferably NTCP proteins derived from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic hamsters, ground shrews, bats, woolly monkeys, capuchins, squirrel monkeys, and ducks. Even more preferably, the NTCP protein is human (e.g., as shown in SEQ ID NO: 7), chimpanzee, or tree shrew NTCP protein. Even more preferably, the NTCP protein is human NTCP protein (e.g., as shown in SEQ ID NO: 7).

[0019] In one or more embodiments, the cells transfected with the exogenous SLCO gene and / or expressing the OATP protein include, but are not limited to, human liver cell lines, mouse liver cell lines, rat liver cell lines, and primary liver cells or liver cell lines of other animal origin. Human liver cell lines include, but are not limited to, HepG2, Huh7, Hep RG, and other human liver-derived cell lines. Mouse-derived liver cell lines include, but are not limited to, mouse-derived liver cells such as Hepa1-6 and Hep56.1D. Rat-derived liver cell lines include, but are not limited to, rat-derived liver cells such as BRL 3A and TC5123. Preferably, the cells are human liver cell lines HepG2, Huh7, Hep RG, mouse liver cell lines Hepa1-6, Hep56.1D, or rat liver cell lines TC5123. Further preferably, the cells are human liver cell lines HepG2 or Huh7.

[0020] In one or more embodiments, the animals transfected with exogenous SLCO genes and / or expressing OATP proteins include, but are not limited to, non-human mammals, birds, and other animals. Non-human mammals include, but are not limited to, mice, rats, dogs, monkeys, tree shrews, pigs, rabbits, marmots, tree shrews, and other mammals. Birds include, but are not limited to ducks, herons, and other birds. Other animals include bats and other animals. The animals are preferably mice, rats, dogs, and monkeys. More preferably, the animals are mice and rats.

[0021] The present invention also includes drugs and lead compounds for treating and / or preventing HBV and / or HDV infection and related diseases obtained by screening using the above-mentioned HBV and / or HDV susceptible cells and / or animal models. Such drugs and lead compounds include, but are not limited to, small molecule drugs, proteins, peptides, antibodies, polyRNAs, polyDNAs, and other drugs. Such drugs and lead compounds for treating and / or preventing HBV and / or HDV infection and related diseases include, but are not limited to, drugs for blocking or inhibiting HBV and / or HDV infection, blocking or inhibiting HBV and / or HDV replication, clearing HBV and / or HDV infection, blocking or inhibiting HBV and / or HDV release, and activating and / or modulating HBV and / or HDV-specific immune responses.

[0022] The present invention also provides a method and a carrier for drug delivery by targeting OATP, wherein the carrier at least retains the biological activity of binding to OATP and delivering the carried drug into cells.

[0023] In one or more embodiments, the OATP includes but is not limited to mammalian, avian, fish, and amphibian OATPs, preferably mammalian and avian OATPs, further preferably mammalian and avian OATP1B subfamily, including OATP1B3 and / or OATP1B1 and / or OATP1B2. More preferably, the OATP is mammalian OATP1B3 and / or OATP1B1 and / or OATP1B2, further preferably OATP1B3 and / or OATP1B1 and / or OATP1B2 derived from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchin monkeys, and squirrel monkeys. Even more preferably, the OATP is human OATP1B3 and / or human OATP1B1 (e.g., as shown in SEQ ID NOs: 1 and 4).

[0024] In one or more embodiments, the vector can bind to an OATP alone or simultaneously to an OATP and NTCP. The NTCP includes, but is not limited to, mammalian, avian, fish, and amphibian NTCP, preferably mammalian and avian NTCP, and more preferably NTCP proteins derived from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchins, squirrel monkeys, and ducks. Even more preferably, the NTCP protein is human, chimpanzee, or tree shrew NTCP protein. Even more preferably, the NTCP protein is human NTCP protein.

[0025] In one or more embodiments, the backbone of the carrier includes, but is not limited to, polypeptides, small molecule drugs, proteins, antibodies, nucleic acids, and other forms, and the backbone can bind to OATP and deliver the carried drug or group into cells and / or organs. Preferably, the backbone can bind to OATP1B3 and / or OATP1B1, and can deliver the carried drug or group into hepatocytes and / or the liver; further preferably, the backbone can bind to human OATP1B3 and / or human OATP1B1, and can deliver the carried drug or group into human hepatocytes and / or the liver.

[0026] In one or more embodiments, the drug or group carried by the carrier includes but is not limited to small molecule drugs, nucleic acids, proteins, antibodies, polypeptides or other drugs or groups, and the drug or group can be linked to the rest of the carrier (skeleton) by covalent bonds and / or non-covalent bonds. In the OATP targeted delivery vector, the drug or group can be directly linked to the rest of the carrier or can be linked through a linker. The small molecule drugs include but are not limited to fluorescent groups, toxins, chemotherapeutics, signal regulatory molecules, cell signal agonists, cell signal inhibitors and other small molecule drugs or groups. The nucleic acids include but are not limited to DNA, RNA and other nucleic acid molecules, and the nucleic acids preferably carried are protein-coding DNA or RNA, antisense nucleic acids, nucleic acids for RNA interference, nucleic acids for gene editing, and viral vectors carrying these nucleotide sequences.

[0027] In one or more embodiments, the methods and vectors are used to deliver drugs or groups including but not limited to mammals, birds, fish and amphibians, preferably mammals and birds, further preferably humans, chimpanzees, bonobos, tree shrews, ducks, marmots, Arctic shrews, shrews, bats, woolly monkeys, capuchins and squirrel monkeys, and even more preferably humans.

[0028] In one or more embodiments, in the methods and carriers, the systems into which the delivered drug or group enters include but are not limited to the digestive system, respiratory system, circulatory system, nervous system, locomotor system, urinary system, reproductive system, endocrine system and other systems; the organs and tissues into which the delivered drug or group enters include but are not limited to the liver, kidney, lung, heart, muscle, fat, brain, nerve, intestine, pancreas and other organs or tissues; the cells into which the delivered drug or group enters include but are not limited to hepatocytes, lung cells, kidney cells, nerve cells, muscle cells, fat cells, blood cells, lymphocytes and other cells. The delivered drug or group preferably enters the digestive system, the delivered drug or group preferably enters the liver, and the delivered drug or group preferably enters hepatocytes.

[0029] In some embodiments of the present invention, the backbone of the OATP targeted delivery vector provided by the present invention includes but is not limited to an OATP targeted delivery polypeptide derived from a surface antigen of a hepadnavirus, which at least retains the biological activity of binding to OATP and delivering the carried drug or group into the cell. The OATP targeted delivery polypeptide derived from the surface antigen of a hepadnavirus is preferably derived from the surface antigen of a protohepadnavirus or an Avihepadnavirus. The OATP targeted delivery polypeptide is further preferably derived from human hepatitis B virus (HBV), tree shrew HBV, woolly monkey hepatitis B virus (WMHBV), woodchuck hepatitis B virus (WHV), duck hepatitis B virus (DHBV), bat HBV and other hepatitis B virus surface antigens. Still further preferably, the polypeptide is derived from the human HBVPre-S1 region, at least retaining the biological activity of binding to OATP and delivering the carried drug or group into the cell.

[0030] In some embodiments of the present invention, the OATP targeted delivery vector derived from the surface antigen of the hepadnavirus is a polypeptide having the following structure:

[0031] Wherein G is glycine, the core sequence is an amino acid sequence derived from the surface antigen of a hepadnavirus, X is an amino acid residue having two amino groups or two carboxyl groups or other groups available for linking reactions, R1 is an N-terminal modification, R2 is absent or is a flanking sequence or other group, and R3 is a carried pharmacophore or a linking group that can interact with a drug or group.

[0032] In one or more embodiments, the core sequence is preferably derived from the surface antigen of the protohepadnavirus or Avihepadnavirus, further preferably from human hepatitis B virus (HBV), tree shrew HBV, woolly monkey hepatitis B virus (WMHBV), woodchuck hepatitis B virus (WHV), duck hepatitis B virus (DHBV), bat HBV and other hepatitis B virus surface antigens and variants thereof, further preferably from the Pre-S1 region of the human HBV surface antigen and variants thereof, and still further preferably from amino acids 14-48 of the Pre-S1 region of the C genotype human HBV surface antigen (an example sequence can be seen in the sequence shown in SEQ ID NO: 97) and variants thereof.

[0033] In one or more embodiments, R1 is preferably an N-terminal essential modification group, which can be selected from a hydrophobic group or other groups. The hydrophobic group can be selected from but not limited to myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, cholesterol, polyethylene glycol, arachidonic acid and other hydrophobic groups; further preferably a fatty acid group, which can be selected from but not limited to myristic acid, palmitic acid, stearic acid and other fatty acids; further preferably myristic acid, palmitic acid, stearic acid groups, and even more preferably myristic acid groups.

[0034] In one or more embodiments, R2 is absent or is a flanking sequence. R2 may or may not have a C-terminal modification. Further preferably, R2 is a flanking sequence derived from the Pre-S1 region of the human HBV surface antigen, with a C-terminal stabilizing modification; even more preferably, a flanking sequence derived from the Pre-S1 region of the human genotype C HBV surface antigen (an example sequence can be found in the amino acid sequence set forth in SEQ ID NO: 141) and variants thereof, with a C-terminal amidation modification.

[0035] In one or more embodiments, R3 is a pharmacophore, or a linking group that can interact with a drug or group. The pharmacophore or linking group is directly or indirectly linked to the backbone of the carrier via covalent and / or non-covalent bonds or other means. There may or may not be a linker between the pharmacophore or linking group and the backbone of the carrier.

[0036] In one or more embodiments, the pharmacophore, drug or group carried by the delivery vector shown in formula (I) includes but is not limited to small molecule drugs, nucleic acids, proteins, antibodies, polypeptides, viruses or other drugs or groups. In some embodiments, R3 is selected from but is not limited to small molecule groups or linking groups that can interact with them, and can be but is not limited to fluorescent groups, toxins, chemotherapeutic drugs, signal regulatory molecules, cell signal agonists, cell signal inhibitors and other small molecule drugs or groups, or linking groups that can interact with them. In some embodiments, R3 is preferably but not limited to fluorescent groups. In some embodiments, R3 is selected from but is not limited to nucleic acids, and can be but is not limited to DNA, RNA and other nucleic acid molecules or linking groups that can interact with them, and its function is to introduce exogenous genes, protein coding, upregulate or downregulate target gene expression; preferably, R3 is selected from but is not limited to protein-coding DNA or RNA, antisense nucleic acids, nucleic acids for RNA interference, nucleic acids for gene editing, or linking groups that can interact with them. In some embodiments of the present invention, the nucleic acid carried by the vector also includes viral vectors carrying these nucleic acid sequences, or linking groups that can interact with them. In some embodiments of the present invention, R3 can be selected from but not limited to small molecule pharmacophores with cell regulatory activity, active proteins, nucleic acids encoding active proteins, and biologically active nucleic acids such as interfering or antisense active protein mRNA, gene encoding nucleic acids and viral vectors, or linking groups that can interact with the small molecule pharmacophores, active proteins, encoding nucleic acids or biologically active nucleic acids.

[0037] In some preferred embodiments of the present invention, the core sequence of the OATP targeted delivery vector derived from the hepadnavirus surface antigen shown in formula (I) is derived from amino acids 14-48 of the Pre-S1 region of the C genotype human HBV surface antigen or a variant thereof; R1 is preferably a myristic acid, palmitic acid, stearic acid, cholesterol, or polyethylene glycol group; the flanking sequence of R2 is preferably derived from amino acids 50-59 of the Pre-S1 region of the C genotype human HBV surface antigen, and its C-terminus is amidated; preferably, the amino acid sequence of the vector is as shown in any one of SEQ ID NOs: 159-164. In some more preferred embodiments of the present invention, the core sequence of the OATP targeted delivery polypeptide vector derived from the hepadnavirus surface antigen shown in formula (I) is preferably derived from amino acids 14-56 of the Pre-S1 region of the C genotype human HBV surface antigen or a variant thereof; R1 is preferably a myristic acid, palmitic acid, stearic acid, cholesterol, or polyethylene glycol group; the R2 flanking sequence is preferably derived from amino acids 58-59 of the Pre-S1 region of the C genotype human HBV surface antigen, and its C-terminus is amidated; preferably, the amino acid sequence of the vector is as shown in SEQ ID NO: 165. In some further preferred embodiments of the present invention, the core sequence of the OATP targeted delivery polypeptide vector derived from the surface antigen of hepadnavirus as shown in formula (I) is preferably derived from amino acids 14-48 of the Pre-S1 region of the surface antigen of human HBV genotype C or a variant thereof; R1 is preferably a myristic acid group; the R2 flanking sequence is preferably derived from amino acids 50-59 of the Pre-S1 region of the surface antigen of human HBV genotype C, with an amidation modification at the C-terminus; and R3 is preferably a small molecule pharmacophore; preferably, the amino acid sequence of the vector is as shown in SEQ ID NO: 159. In one embodiment of the present invention, the core sequence of the OATP targeted delivery polypeptide vector derived from the surface antigen of hepadnavirus as shown in formula (I) is selected from amino acids 14-48 of the Pre-S1 region of the surface antigen of human HBV genotype C; R1 is selected from a myristic acid group; the R2 flanking sequence is selected from amino acids 50-59 of the Pre-S1 region of the surface antigen of human HBV genotype C, with an amidation modification at the C-terminus; and R3 is selected from a FITC fluorescent group.

[0038] The present invention also provides a hepadnavirus surface antigen-derived peptide that binds to an OATP and inhibits its transport function. This peptide inhibits the transport function of the OATP by binding to the OATP. The OATP includes, but is not limited to, mammalian, avian, fish, and amphibian OATPs, preferably mammalian and avian OATPs, and more preferably, the mammalian and avian OATP1B subfamily, including OATP1B3 and / or OATP1B1 and / or OATP1B2. More preferably, the OATP is mammalian OATP1B3 and / or OATP1B1 and / or OATP1B2, and even more preferably, OATP1B3 and / or OATP1B1 and / or OATP1B2 derived from humans, chimpanzees, bonobos, tree shrews, marmots, arctic ground shrews, ground shrews, bats, woolly monkeys, capuchin monkeys, and squirrel monkeys. Even more preferably, the OATP is human OATP1B3 and / or human OATP1B1.

[0039] Preferably, the amino acid sequence of the polypeptide is derived from a hepadnavirus surface antigen. Preferably, the amino acid sequence of the polypeptide is derived from an HBV viral surface antigen, including but not limited to human hepatitis B virus (HBV), tree shrew HBV, woolly monkey hepatitis B virus (WMHBV), woodchuck hepatitis B virus (WHV), duck hepatitis B virus (DHBV), bat HBV and other hepatitis B viruses. More preferably, the amino acid sequence of the polypeptide is derived from a human hepatitis B virus (HBV) surface antigen (HBsAg). The human hepatitis B virus (HBV) includes but is not limited to genotypes A, B, C, D, E, F, G, H, and I HBV. Further preferably, the amino acid sequence of the polypeptide is derived from the human hepatitis B virus (HBV) surface antigen (HBsAg) Pre-S1 region, further preferably from amino acids 2-77 (D genotype) or amino acids 13-88 (C genotype) of the human hepatitis B virus (HBV) surface antigen (HBsAg) Pre-S1 region, more preferably from amino acids 2-48 (D genotype) or amino acids 13-59 (C genotype) of the HBsAg Pre-S1 region, more preferably from amino acids 13-59 (C genotype) of the HBsAg Pre-S1 region. Preferably, the N-terminus of the polypeptide is modified with a fatty acid or other modification, further preferably with myristic acid. More preferably, the amino acid sequence of the polypeptide is as shown in SEQ ID NO: 67 or 68, and the N-terminus of the polypeptide is modified with myristic acid.

[0040] Preferably, the transport function of the OATP includes but is not limited to the transport of statins, sartans, tinibs, anti-tumor drugs such as platinum drugs and paclitaxel drugs and other drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1: Map of recombinant lentivirus WL1142. In the figure, hHEPALK3 represents hOATP1B3.

[0042] Figure 2: Map of the recombinant lentivirus WL1143. hGBIO represents hNTCP.

[0043] Figure 3: Structure of L47-FITC (A), HPLC identification diagram (B) and mass spectrometry identification diagram (C).

[0044] Figure 4: HPLC identification chart (A) and mass spectrometry identification chart (B) of L47.

[0045] Figure 5: Hela cells incubated with PBS served as negative controls for FITC and PI staining; Hela cells fixed with paraformaldehyde and incubated with L47-FITC and PI, respectively, served as positive controls for FITC and PI staining, and the resulting signal fluorescence intensities were measured.

[0046] Figure 6: Flow cytometry results of the BLANK group.

[0047] Figure 7: Flow cytometry results of the single staining group.

[0048] Figure 8: Flow cytometry results of the double-staining group.

[0049] Figure 9: Comparison of FITC signals in Hela cells stained with blank PBS, Hela cells stained with L47-FITC / PI, Hela cells, Hela-hNTCP, Hela-hOATP1B3, and Hela-hNTCP / hOATP1B3 cells.

[0050] Figure 10: Fluorescence intensity of Hela cells stained with blank PBS, Hela cells double-stained with L47-FITC / PI, Hela cells, Hela-hNTCP, Hela-hOATP1B3, and Hela-hNTCP / hOATP1B3 cells.

[0051] Figure 11: Fluorescence signals of different cells after administration of PBS or L47-FITC under Leica LED8 cyan 475±14nm excitation light.

[0052] FIG12 : Inhibitory effect of L47 on the uptake of E217βG by OATP1B1.

[0053] FIG13 : Inhibitory effect of L47 on the uptake of E217βG by OATP1B3.

[0054] FIG14 : Inhibitory effect of hepalatide on the uptake of valsartan by OATP1B1.

[0055] FIG15 : Inhibitory effect of hepalatide on the uptake of valsartan by OATP1B3.

[0056] FIG16 : Inhibitory effect of hepalatide on the uptake of atorvastatin by OATP1B1.

[0057] Figure 17: Map of recombinant lentivirus WL1281. In the figure, both hHEPA LK1 and WL1281 express hOATP1B1.

[0058] Figure 18: HBsAg and HBeAg concentrations in the culture supernatant on day 9 after infection in Experiment 1 of Example 6.

[0059] Figure 19: HBsAg and HBeAg concentrations in the culture supernatant on day 9 after infection in Experiment 2 of Example 6. DETAILED DESCRIPTION

[0060] The present invention found that both human NTCP and human OATP1B3 can bind to polypeptides derived from HBVPre-S1, but the binding of hNTCP to HBVPre-S1-derived polypeptides is limited to the cell membrane and does not mediate the entry of the polypeptide into the cell. However, the binding of HBVPre-S1-derived polypeptides to OATP1B3 can mediate the entry of the polypeptide into the cell. In addition, co-expression of OATP1B3 and NTCP can enhance the binding of HBVPre-S1-derived polypeptides to cells. It is well known in the art that the HBVPre-S1 region is a key region for mediating HBV infection, and this region binds to the HBV infection functional receptor NTCP [Yan H et al. Elife. 2012; 1: e00049.; Ni Y et al. Gastroenterology. 2014; 146(4): 1070-83.]; OATP1B3 has a high homology with OATP1B1 [ J, et al. J Biol Chem. 2000; 275(30): 23161-8.], and both can form heterodimers with NTCP [Zhang Y, et al. PLoS One. 2017; 12(6): e0180257.]. Combining the findings of the present invention with the known knowledge in the art, it is reasonable to infer that: 1) human OATP1B3 is a receptor for HBV infection, and HBV can bind to human OATP1B3 or human OATP1B1 through its Pre-S1 region, mediating HBV entry into cells; 2) human OATP1B3 or human OATP1B1 forms a co-receptor for HBV infection with NTCP. Given that other animals also have hepadnavirus infections and homologous OATP molecules, this infection mechanism can be extended to hepadnavirus infection of other animals.

[0061] Therefore, this paper proposes:

[0062] 1) By preventing or reducing the interaction between HBV and / or HDV and OATP, or preventing or reducing the expression and / or function of OATP, infections and related diseases caused by HBV and / or HDV can be treated and / or prevented;

[0063] 2) Cells or animals that are not susceptible or not easily infected by HBV or HDV can be transfected with the exogenous SLCO gene, or with both the SLCO and SLC10A1 genes, to express exogenous OATP proteins, or to co-express OATP and NTCP proteins, thereby generating cell and animal models susceptible to HBV and / or HDV. These cell and animal models can be used to screen for drugs or lead compounds for the treatment and / or prevention of HBV and / or HDV infection.

[0064] 3) Drugs, exogenous genes, and expression-regulating substances can be delivered into cells by targeting OATP for the prevention and / or treatment of related diseases;

[0065] 4) By binding HBVPre-S1-derived peptides to OATP, the OATP transport function is inhibited, and the pharmacokinetics of drugs that use OATP as a transporter are regulated to improve the efficacy and safety of these drugs.

[0066] Terms and Definitions

[0067] It should be understood that within the scope of the present invention, the above-mentioned various technical features of the present invention and the various technical features specifically described below (such as in the examples) can be combined with each other to form a preferred technical solution. Unless otherwise specified, all technical and scientific terms used herein are consistent with the meanings commonly understood by those of ordinary skill in the art to which this disclosure belongs. For the purpose of this disclosure, the following terms are defined as follows. The article "one" refers to one or more (i.e., at least one) grammatical objects of the article. For example, "an element" refers to one element or more than one element. Unless otherwise clearly stated in the text, the term "or" is used interchangeably with the term "and / or". With respect to the scope of the terms "containing", "including", "having" or their grammatical variants used in this disclosure or claims, these terms can be used in a similar manner to the term "comprising" when used as a transition word in the claims. The term "including" or its grammatical variants means "including but not limited to" and can be used interchangeably with "including but not limited to". The term "about" refers to an amount, level, value, numerical value, frequency, percentage, dimension, size, quantity, weight, or length that varies by approximately 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% compared to a reference amount, level, value, numerical value, frequency, percentage, dimension, size, quantity, weight, or length. When the term "about" is used in conjunction with a numerical range, the term modifies the range, extending its limits above or below the numerical value. Generally, the term "about," when modifying a numerical value, means within 10% above and below the numerical value.

[0068] As used herein, the terms "polypeptide," "peptide," and "protein" are used interchangeably and include full-length proteins and fragments thereof, as well as variants of the full-length proteins and fragments thereof.

[0069] As used herein, "variant" related to the polypeptides described herein refers to a polypeptide that differs from a given polypeptide in amino acid sequence but retains one or more biological activities described herein of the given polypeptide. The biological functions described herein include acting with OATP, binding to OATP, delivering drugs into cells, and inhibiting OATP transport function. The polypeptide variants described herein may have one or more amino acid additions (such as insertions), deletions, or substitutions relative to a given polypeptide. In some embodiments, the polypeptide variants described herein may have 1-30, 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid additions (such as insertions), deletions, or substitutions relative to a given polypeptide. For example, the amino acid sequence of a variant may contain conservative substitutions of amino acids. Conservative substitutions of amino acids, i.e., replacing an amino acid with a different amino acid having similar properties (such as hydrophilicity and the distribution of charged regions),

[0070] The terms "conservative amino acid substitution" and "conservative substitution" are used interchangeably herein to refer to a specified amino acid exchange within a group of amino acids, wherein an amino acid is exchanged for a different amino acid of similar size, structure, charge and / or polarity. Because conservative substitutions generally involve minor changes, they do not significantly alter the biological activity of the polypeptide. These minor changes can be identified, in part, by considering the hydropathic index of the amino acids based on the hydrophobicity and charge of the amino acids. Amino acids with similar hydropathic index and hydropathicity values ​​can be substituted for each other and still retain protein function. The hydropathic index and hydropathicity values ​​of an amino acid are influenced by the specific side chain of the amino acid. Consistent with this observation, amino acid substitutions that match biological function rely on the relative similarity of the amino acids, particularly those of their side chains, as manifested in hydrophobicity, hydrophilicity, charge, size and other properties.

[0071] Families of amino acid residues with similar side chains are well known in the art and include basic side chains (e.g., leucine, arginine, and histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, in some embodiments, an amino acid residue in a polypeptide can be replaced with another amino acid residue from the same side chain family. In other embodiments, an amino acid sequence can be replaced with an amino acid sequence that is structurally similar but differs in the order and / or composition of the side chain family members. In other embodiments, mutations can be introduced randomly throughout the entire sequence or a portion of the polypeptide. Examples of conservative amino acid substitutions include, for example, replacing one of the aliphatic or hydrophobic amino acids Ala, Val, Leu, and Ile with another amino acid in the group of four amino acids; replacement between hydroxyl-containing residues Ser and Thr; replacement between acidic residues Asp and Glu; replacement between amide residues Asn and Gln; replacement between basic residues Lys, Arg, and His; replacement between aromatic residues Phe, Tyr, and Trp; and replacement between small amino acids Ala, Ser, Thr, Met, and Gly. It is reasonably expected that conservative substitutions, such as replacing a conservative amino acid with a similar, structurally related amino acid, will not have a substantial effect on the biological activity of the polypeptide.

[0072] "Variants" of polypeptides also include polypeptides that have a certain degree of identity with a given polypeptide, such as polypeptides that are at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a given polypeptide. As used herein, "variants" of polypeptides also include polypeptides that contain a portion of a given polypeptide corresponding to the native sequence of the protein. "Variants" may also refer to fusion proteins or chimeric proteins, which are polypeptides that are derived from two or more different sources. Non-limiting examples of fusion proteins described herein include, for example, a fusion protein of a polypeptide derived from a hepadnavirus with another polypeptide derived from a non-hepadnavirus protein, a fusion protein of two polypeptides derived from different hepadnavirus genotypes, and a fusion protein of two polypeptides derived from different regions of a surface antigen of any one hepadnavirus genotype or two polypeptides derived from different sequences within a region of a surface antigen of any one hepadnavirus genotype.

[0073] The term "sequence identity" (e.g., 50% identity to a sequence) refers to the degree to which sequences are identical when compared amino acid to amino acid over a comparison window. In some embodiments, the polypeptides described herein may contain an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequence of a given polypeptide and still retains one or more biological activities of the given polypeptide. "Percent identity" or "% identity" can be calculated by optimally aligning the two sequences over a comparison window, determining the number of positions at which the identical amino acid appears in the two sequences to obtain the number of matching positions, and dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal sequence alignment for alignment over a comparison window can be performed by using a computer to execute algorithms known in the art, such as The program of the family, or by visual observation, adopts any method selected to produce the best arrangement (that is, to produce the highest homology percentage in the comparison window). For sequence comparison, a sequence is used as a reference sequence, and the test sequence is compared with the reference sequence. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, and if necessary, subsequence coordinates can be designed and sequence algorithm program parameters are set. Then, the sequence comparison algorithm calculates the current sequence identity of the test sequence relative to the reference sequence based on the set program parameters. The setting of sequence algorithm program parameters is well known in the art. For example, the comparison window can be set to cover the entire length of any one or two comparison sequences, such as covering the entire length of the reference sequence, while allowing the presence of a difference of up to 5% of the total amino acid number of the reference sequence.

[0074] The term "variant" also includes polypeptides that have the same amino acid sequence as a given polypeptide, retain one or more biological activities of the given polypeptide, but have been chemically and / or post-translationally modified in a manner different from that of the given polypeptide. "Variant" can also be used to describe polypeptides that have been differentially processed, such as by proteolysis, phosphorylation, or other post-translational modifications, but retain one or more biological activities of the given polypeptide. "Variant" can include natural and / or non-natural amino acid residues. These terms also include post-translationally modified polypeptides, including, for example, glycosylated, sialylated, acetylated, and / or phosphorylated proteins. The term "variant" also includes polypeptides that have been chemically modified at one or more amino acid residues.

[0075] As used herein, unless otherwise indicated, a "variant" of a polypeptide includes fragments of the variant.

[0076] The term "fragment" refers to a polypeptide consisting of only a contiguous portion of the intact full-length polypeptide sequence and structure. Such fragments may include C-terminal and / or N-terminal deletions of the native polypeptide. Fragments typically possess certain functions of the full-length sequence. Depending on the full-length sequence, the length of a fragment possessing the desired function may vary, for example, typically being at least 10 contiguous amino acid residues in length.

[0077] The term "coding sequence" or a sequence "encoding" a selected polypeptide refers to a nucleic acid molecule that is transcribed (for DNA) and translated (for mRNA) into a polypeptide in vitro or in vivo when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence are determined by a start codon at the 5' (amino) terminus and a translation stop codon at the 3' (carboxyl) terminus. A transcription termination sequence may be located 3' to the coding sequence.

[0078] The term "variant" as used herein in connection with a gene or coding sequence includes, but is not limited to, changes in the sequence produced by base changes in a DNA or RNA encoding nucleic acid sequence, such as base substitutions, insertions, deletions, or duplications. The terms "nucleic acid," "nucleotide," and "base" are used interchangeably herein. As used herein, a "variant" related to a coding sequence described herein refers to a variant that differs from a given coding sequence in base sequence but retains one or more biological activities of the expressed protein, including a full coding sequence and its fragments, as well as a full-length coding sequence and its fragments. Unless otherwise indicated herein, a "variant" as used herein in connection with a gene or coding sequence includes fragments of the variant. The variant coding sequences described herein may have one or more base additions (such as insertions), deletions, or substitutions relative to a given nucleic acid sequence. In some embodiments, the variant coding sequences described herein may have 1-100, 1-50, 1-20, 1-10, 1-5, or 1-3 (including all integers within these ranges) base additions (such as insertions), deletions, or substitutions relative to a given coding sequence. For example, a coding sequence may contain a synonymous mutation of a base. Due to the degeneracy of biological codons, after a base in the coding sequence is changed, the same amino acid is translated at the original amino acid position. The base substitution does not cause an amino acid change, which is also called a neutral mutation or silent mutation. For example, UCU, UCC, UCA, UCG, AGU, and AGC can all encode serine Ser, and the codes of these serines can be interchangeable. For example, replacing a codon encoding an amino acid with a codon encoding a different amino acid with similar properties (such as hydrophilicity and charge level and distribution of charged regions) can achieve conservative amino acid substitutions in the expressed protein without significantly changing the biological activity of the expressed protein. Families of amino acid residues with similar side chains are well known in the art and include basic side chains (e.g., leucine, arginine, and histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, in some embodiments, a codon encoding an amino acid in a protein in a coding sequence can be replaced by a codon encoding another amino acid of the same side chain family. For example, the codons UCU, UCC, UCA, UCG, AGU, AGC encoding serine in a coding sequence can be replaced by the codons ACU, ACC, ACA, ACG encoding threonine.Examples of conservative amino acid codon substitutions include, for example, replacing one amino acid codon from the group of four amino acids with one codon for the aliphatic or hydrophobic amino acids Ala, Val, Leu, and Ile; substitutions between codons for hydroxyl residues Ser and Thr; substitutions between codons for acidic residues Asp and Glu; substitutions between codons for amide residues Asn and Gln; substitutions between codons for basic residues Lys, Arg, and His; substitutions between codons for aromatic residues Phe, Tyr, and Trp; and substitutions between codons for small amino acids Ala, Ser, Thr, Met, and Gly. It is reasonably foreseeable that conservative amino acid codon substitutions, such as replacing a conservative amino acid codon with a codon for a similar, structurally related amino acid, will not substantially affect the biological activity of the protein expressed by the coding sequence.

[0079] The term "variant" also includes deletions or insertions of one or more bases in a given coding sequence, particularly deletions or insertions of three bases, such as deletions or insertions of 3, 6, 9, 15, 30, 60, 90, or 300 bases, without causing changes in a series of downstream codons, changes in the reading frame, or frame shift mutations.

[0080] The term "variant" also includes various chemical modifications of DNA, RNA bases, and ribose sugars of a given coding sequence and its variants. These include, but are not limited to, phosphorylation, methylation, PEGylation, histone modification, and the like. It also encompasses conjugates of a given coding sequence and its variants with proteins, polypeptides, and other chemical molecules. It also encompasses capping of a given coding sequence and its variants, such as with an m7GTP cap, or tailing of a given coding sequence and its variants, such as with a poly-A tail.

[0081] The term "variant" also includes variants that have a certain degree of identity to a given coding sequence, such as at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to a given coding sequence. The term "sequence identity" (e.g., 50% identity to a sequence) refers to the degree to which sequences are identical when compared base for base over a comparison window. In some embodiments, the coding sequences described herein may contain a base sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the base sequence of a given coding sequence and still retain one or more biological activities of the protein expressed by the given coding sequence. "Percent identity" or "% identity" can be calculated by optimally aligning the two sequences over a comparison window, determining the number of positions where the identical base occurs in the two sequences to obtain the number of matched positions, and dividing the number of matched positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal sequence alignment for alignment over a comparison window can be performed by using a computer to execute algorithms known in the art, such as The program of the family, or by visual observation, adopts any method selected to produce the best arrangement (that is, to produce the highest homology percentage in the comparison window). For sequence comparison, a sequence is used as a reference sequence, and the test sequence is compared with the reference sequence. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, and if necessary, subsequence coordinates can be designed, and the sequence algorithm program parameters are set. Then, the sequence comparison algorithm calculates the current sequence identity of the test sequence relative to the reference sequence based on the set program parameters. The setting of the sequence algorithm program parameters is well known in the art. For example, the comparison window can be set to cover the entire length of any one or two comparison sequences, such as covering the entire length of the reference sequence, while allowing the presence of a difference of up to 5% of the total base number of the reference sequence.

[0082] The term "transformation" as used herein refers to the insertion of an exogenous polynucleotide into a host cell, regardless of the method of insertion: for example, transformation by direct uptake, transfection, infection, etc. Specific transfection methods are further described below. The exogenous polynucleotide can be maintained as an unintegrated vector, such as an episome, or can be integrated into the host genome.

[0083] The term "host cell" is a cell that has been transformed, or is capable of being transformed, by an exogenous DNA sequence.

[0084] Methods and agents for treating or preventing HBV and / or HDV infection and related diseases

[0085] Provided herein are methods and agents for preventing and / or treating hepadnavirus and / or HDV infection or diseases associated with such infection in animals by preventing or reducing the interaction between hepadnavirus and / or HDV and OATPs, and / or preventing or reducing the expression and / or function of OATPs. Also provided herein are methods and agents for preventing and / or treating hepadnavirus and / or HDV infection or diseases associated with such infection in animals by preventing or reducing the interaction between hepadnavirus and / or HDV and OATPs, and / or preventing or reducing the expression and / or function of OATPs. Furthermore, provided herein are substances for preventing and / or treating hepadnavirus and / or HDV infection or diseases associated with such infection in animals that can prevent or reduce the interaction between hepadnavirus and / or HDV and OATPs, and / or preventing or reducing the expression and / or function of OATPs.

[0086] Herein, the animals include but are not limited to mammals, birds, fishes, amphibians and other animals.

[0087] The mammals include, but are not limited to, primates, tree shrews, rodents, bats, even-toed ungulates, Carnivora, and other mammals. The birds include, but are not limited to, ducks, herons, and other avian animals. The primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. The hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other humanoids. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, Macaca fascicularis and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other myomorphs. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora. The animals described herein particularly include humans, primates, tree shrews, rodents, birds, and bats, and in particular, include humans, chimpanzees, bonobos, tree shrews, ducks, marmots, arctic ground squirrels, ground shrews, bats, woolly monkeys, capuchin monkeys, and squirrel monkeys. In particularly preferred embodiments, the animals described herein are humans.

[0088] The hepadnaviruses include, but are not limited to, orthohepadnavirus (OHV), avihepadnavirus, metahepadnavirus, herpetohepadnavirus, and other hepadnaviruses. The OHVs include, but are not limited to, orthohepadnaviruses that infect humans (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkeys, woodchuck hepatitis virus (WHV), arctic ground shrews, ground shrews (GSHV), tree shrews, chimpanzees, bats (BHV), and other animals. Exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers AF046996 (WMHBV), NC_043528 (CMHBV), NC_004107 (WHV), NC_001484 (GSHV), and JX941468 (BHV). The Avihepadnavirus includes but is not limited to Avihepadnavirus that infects ducks (duck HBV, DHBV), herons (heron HBV, HHBV) and other birds. Exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers NC_001344 (DHBV) and NC_001486 (HHBV).

[0089] The original hepadnavirus (HBV) that infects humans described herein includes, but is not limited to, genotypes A, B, C, D, E, F, G, H and I genotype HBV, and exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G) and AB179747 (genotype H).

[0090] The HDVs described herein include, but are not limited to, genotypes 1, 2, 3, 4, 5, 6, 7, and 8 HDVs, and exemplary genomic sequences can be found in, but are not limited to, GenBank accession numbers MG711803 (genotype 1), MK234593 (genotype 2), LT604954 (genotype 3), MK248835 (genotype 4), JX888107 (genotype 5), MG711780 (genotype 6), MG711804 (genotype 7), and LT604973 (genotype 8).

[0091] The OATPs described herein include but are not limited to the OATP superfamily of mammals, birds, fishes, amphibians and other animal sources. In some embodiments, the OATP superfamily includes but is not limited to OATP1, OATP2, OATP3, OATP4, OATP5, OATP6, OATP7, OATP8, OATP9, OATP10, OATP11 and other OATP families. In some embodiments, the OATP family includes but is not limited to the OATP family expressed in hepatocytes. In some embodiments, the OATP includes but is not limited to the OATP1B subfamily. In some preferred embodiments, the OATP includes but is not limited to the OATP1B subfamily expressed in hepatocytes. In some preferred embodiments, the OATP includes but is not limited to OATP1B3, OATP1B1, OATP1B2 and other OATP1B subfamily members.

[0092] The OATP1B3 protein and SLCO1B3 gene described herein can be derived from mammals, birds, fish, amphibians, and other animals. Mammals include, but are not limited to, primates, tree shrews, rodents, bats, even-toed ungulates, Carnivora, and other mammals that can be infected with hepadnaviruses, or mammals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Birds include, but are not limited to, ducks, herons, and other avian animals that can be infected with avihepadnaviruses, or avian animals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates that can be infected with hepadnaviruses, or primates whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species that can be infected with hepadnaviruses, or hominoid species whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The New World monkeys include but are not limited to woolly monkeys, capuchin monkeys, squirrel monkeys and other New World monkeys that can be infected with hepadnaviruses, or New World monkeys whose liver cells can be infected with hepadnaviruses after being transfected with exogenous SLC10A1 genes or expressing exogenous NTCP proteins.The Old World monkeys include, but are not limited to, cynomolgus macaques (Macaca fascicularis) and other Old World monkeys that can be infected with hepadnaviruses, or Old World monkeys whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents that can be infected with hepadnaviruses, or rodents whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae that can be infected with hepadnaviruses, or Sciuridae whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs that can be infected with hepadnaviruses, or myomorphs whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The artiodactyls include, but are not limited to, pigs and other even-toed ungulates that can be infected with hepadnaviruses, or even-toed ungulates that can be infected with hepadnaviruses after their liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein. The carnivores include, but are not limited to, dogs and other carnivores that can be infected with hepadnaviruses, or carnivores whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses.

[0093] The OATP1B3 amino acid sequences described herein include, but are not limited to, the OATP1B3 amino acid sequence from human (SEQ ID NO: 1, NCBI Reference Sequence: NM 019844), the OATP1B3 amino acid sequence from chimpanzee (NCBI Reference Sequence: XM_016923821), the OATP1B3 amino acid sequence from tree shrew (NCBI Reference Sequence: XM_006155085), the OATP1B3 amino acid sequence from cynomolgus monkey (NCBI Reference Sequence: NM_001283191), the OATP1B3 amino acid sequence from Arctic ground squirrel (NCBI Reference Sequence: XM_026391932), the OATP1B3 amino acid sequence from rat (NCBI Reference Sequence: XM_032905480), the OATP1B3 amino acid sequence from porcine (NCBI Reference Sequence: XM_006155085), the OATP1B3 amino acid sequence from cynomolgus monkey (NCBI Reference Sequence: NM_001283191), the OATP1B3 amino acid sequence from Arctic ground squirrel (NCBI Reference Sequence: XM_026391932), the OATP1B3 amino acid sequence from rat (NCBI Reference Sequence: XM_032905480), the OATP1B3 amino acid sequence from porcine (NCBI Reference Sequence: XM_006155085), the OATP1B3 amino acid sequence from Sequence: NM_001315607), the OATP1B3 amino acid sequence from dog (NCBI Reference Sequence: NM_001166047), and the OATP1B3 amino acid sequences from the other animals mentioned above.

[0094] The OATP1B3 nucleic acid coding sequence (CDS) described herein includes, but is not limited to, the OATP1B3 nucleic acid coding sequence from human (SEQ ID NO: 2, NCBI Reference Sequence: NM_019844), the OATP1B3 nucleic acid coding sequence from chimpanzee (NCBI Reference Sequence: XM_016923821), the OATP1B3 nucleic acid coding sequence from tree shrew (NCBI Reference Sequence: XM_006155085), the OATP1B3 nucleic acid coding sequence from cynomolgus monkey (NCBI Reference Sequence: NM_001283191), the OATP1B3 nucleic acid coding sequence from Arctic ground squirrel (NCBI Reference Sequence: XM_026391932), the OATP1B3 nucleic acid coding sequence from rat (NCBI Reference Sequence: XM_032905480), the OATP1B3 nucleic acid coding sequence from porcine (NCBI Reference Sequence: XM_006155085), the OATP1B3 nucleic acid coding sequence from cynomolgus monkey (NCBI Reference Sequence: NM_001283191), the OATP1B3 nucleic acid coding sequence from Arctic ground squirrel (NCBI Reference Sequence: XM_026391932), the OATP1B3 nucleic acid coding sequence from rat (NCBI Reference Sequence: XM_032905480), the OATP1B3 nucleic acid coding sequence from porcine (NCBI Reference Sequence: XM_006155085), the OATP1B3 nucleic acid coding sequence from Sequence: NM_001315607), the OATP1B3 nucleic acid coding sequence from dog (NCBI Reference Sequence: NM_001166047), and the OATP1B3 nucleic acid coding sequences from the other animals mentioned above.

[0095] The SLCO1B3 gene sequence described herein includes, but is not limited to, a SLCO1B3 gene sequence derived from humans (SEQ ID NO: 3, NCBI Reference Sequence: NM_019844), a SLCO1B3 gene sequence derived from chimpanzees (genebank Gene ID: 749254), a SLCO1B3 gene sequence derived from tree shrews (genebank GeneID: 102476084), a SLCO1B3 gene sequence derived from cynomolgus monkeys (genebank GeneID: 102136593), a SLCO1B3 gene sequence derived from Arctic ground squirrels (genebank GeneID: 113185010), a SLCO1B3 gene sequence derived from rats (genebank GeneID: 116903127), a SLCO1B3 gene sequence derived from pigs (genebank GeneID: 100620829), and a SLCO1B3 gene sequence derived from dogs (genebank GeneID: 403519).and SLCO1B3 gene sequences from the above-mentioned other animals, including but not limited to gene banks NC_050564, NC_013676, NW_017869963, NW_006501111, NW_022196903, NW_003613944, NW_004801640, NW_004949097, NW_015351262, NW_024404945, NW_020539983, NW_023144717, NW_006711288, NW_019154091, N W_004569192, NC_044309, NW_020339853, NC_018729, NW_017619867, NW_023365217, NC_051831, NW_02065 6135, NW_022631003, NC_045619, NW_004451484, NC_043709, NW_020356453, NC_048233, NW_020312893, NW _020998839, NW_006383153, NW_023270504, NC_032654, NC_010447, NW_006781351, NW_023601653, NW_011 889225, NC_009149, NW_007675154, NW_023458025, NW_006159939, NW_023666069, NW_012009027, NC_0222 82, NC_041764, NW_012010756, NC_044984, NC_037679, NW_012109719, NW_022681456, NW_016809129, NC_044558, NC_045443, NC_044614, NC_048251, NC_036891, NC_000012, NC_036915, NC_044403, NW_022611650, NW_024100944, NC_033666, NW_006408616, and other SLCO1B3 gene sequences.

[0096] Also encompassed herein are variants and homologous sequences of the aforementioned OATP1B3 amino acid sequences, nucleic acid coding sequences, and SLCO1B3 gene sequences.

[0097] The OATP1B1 protein and SLCO1B1 gene described herein can be derived from mammals, birds, fish, amphibians, and other animals. Mammals include, but are not limited to, primates, tree shrews, rodents, bats, even-toed ungulates, Carnivora, and other mammals that can be infected with hepadnaviruses, or mammals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Birds include, but are not limited to, ducks, herons, and other avian animals that can be infected with avihepadnaviruses, or avian animals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates that can be infected with hepadnaviruses, or primates whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species that can be infected with hepadnaviruses, or hominoid species whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The New World monkeys include but are not limited to woolly monkeys, capuchin monkeys, squirrel monkeys and other New World monkeys that can be infected with hepadnaviruses, or New World monkeys whose liver cells can be infected with hepadnaviruses after being transfected with exogenous SLC10A1 genes or expressing exogenous NTCP proteins.The Old World monkeys include, but are not limited to, cynomolgus macaques (Macaca fascicularis) and other Old World monkeys that can be infected with hepadnaviruses, or Old World monkeys whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents that can be infected with hepadnaviruses, or rodents whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae that can be infected with hepadnaviruses, or Sciuridae whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs that can be infected with hepadnaviruses, or myomorphs whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The artiodactyls include, but are not limited to, pigs and other even-toed ungulates that can be infected with hepadnaviruses, or even-toed ungulates that can be infected with hepadnaviruses after their liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein. The carnivores include, but are not limited to, dogs and other carnivores that can be infected with hepadnaviruses, or carnivores whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses.

[0098] The OATP1B1 amino acid sequence described herein includes, but is not limited to, an OATP1B1 amino acid sequence derived from humans (SEQ ID NO: 4, NCBI Reference Sequence: NM_006446), an OATP1B1 amino acid sequence derived from bonobos (NCBI Reference Sequence: XM_003828901), an OATP1B1 amino acid sequence derived from cynomolgus monkeys (NCBI Reference Sequence: NM_001284540), and OATP1B1 amino acid sequences derived from the above-mentioned other animals.

[0099] The OATP1B1 nucleic acid coding sequence described herein includes, but is not limited to, the OATP1B1 nucleic acid coding sequence derived from humans (SEQ ID NO: 5, NCBI Reference Sequence: NM_006446), the OATP1B1 nucleic acid coding sequence derived from bonobos (NCBI Reference Sequence: XM_003828901), the OATP1B1 nucleic acid coding sequence derived from cynomolgus monkeys (NCBI Reference Sequence: NM_001284540), and the OATP1B1 nucleic acid coding sequences derived from the above-mentioned other animals.

[0100] The SLCO1B1 gene sequences described herein include, but are not limited to, the SLCO1B1 gene sequence from humans (SEQ ID NO: 6, Genebank Gene ID: 10599), the SLCO1B1 gene sequence from bonobos (Genebank Gene ID: 100976318), the SLCO1B1 gene sequence from cynomolgus macaques (Genebank GeneID: 102137347), and the SLCO1B1 gene sequences from the above-mentioned other animals, including, but not limited to, the SLCO1B1 gene sequences from genebank NC_000012, NC_041764, NC_036891, NC_022282, NC_048391, NW_012009027, NW_012014011, NC_037679, NW_012109719, NC_044614, NC_048251, NC_036915, NW_006399825, NW_023666069, NW_016809129, NC_044558, NC_044403, NW_022611650, NW_012112030, NC_045443, NW_006399825, and other SLCO1B1 gene sequences.

[0101] The present invention also includes variants and homologous sequences of the above-mentioned OATP1B1 amino acid sequence, nucleic acid coding sequence and SLCO1B1 gene sequence.

[0102] In some animals, there is an OATP1B2 protein that is homologous to OATP1B3, and a SLCO1B2 gene that is homologous to SLCO1B3.

[0103] The OATP1B2 protein and SLCO1B2 gene described herein can be derived from mammals, birds, fish, amphibians, and other animals. The mammals include, but are not limited to, primates, tree shrews (tupaia), rodents, bats, even-toed ungulates (Artiodactyla), Carnivora, and other mammals that can be infected with hepadnavirus, or mammals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnavirus; the birds include, but are not limited to, ducks, herons, and other avian animals that can be infected with avihepadnavirus, or avian animals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnavirus. Primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates that can be infected with hepadnaviruses, or primates whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species that can be infected with hepadnaviruses, or hominoid species whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The New World monkeys include but are not limited to woolly monkeys, capuchin monkeys, squirrel monkeys and other New World monkeys that can be infected with hepadnaviruses, or New World monkeys whose liver cells can be infected with hepadnaviruses after being transfected with exogenous SLC10A1 genes or expressing exogenous NTCP proteins.The Old World monkeys include, but are not limited to, cynomolgus macaques (Macaca fascicularis) and other Old World monkeys that can be infected with hepadnaviruses, or Old World monkeys whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The rodents include, but are not limited to, Sciuridae animals, myomorphs, and other rodents that can be infected with hepadnaviruses, or rodents whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The Sciuridae animals include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae animals that can be infected with hepadnaviruses, or Sciuridae animals whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs that can be infected with hepadnaviruses, or myomorphs whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses. The artiodactyls include, but are not limited to, pigs and other even-toed ungulates that can be infected with hepadnaviruses, or even-toed ungulates that can be infected with hepadnaviruses after their liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein. The carnivores include, but are not limited to, dogs and other carnivores that can be infected with hepadnaviruses, or carnivores whose liver cells are transfected with an exogenous SLC10A1 gene or express an exogenous NTCP protein and can be infected with hepadnaviruses.

[0104] The OATP1B2 amino acid sequence described herein includes, but is not limited to, the OATP1B2 amino acid sequence from rat (NCBI Reference Sequence: NM_031650), the OATP1B2 amino acid sequence from mouse (NCBI Reference Sequence: NM_020495), and the OATP1B2 amino acid sequence from the above-mentioned other animals.

[0105] The OATP1B2 nucleic acid coding sequence described herein includes, but is not limited to, the OATP1B2 nucleic acid coding sequence derived from rat (NCBI Reference Sequence: NM_031650), the OATP1B2 nucleic acid coding sequence derived from mouse (NCBI Reference Sequence: NM_020495), and the OATP1B2 nucleic acid coding sequence derived from the above-mentioned other animals.

[0106] The SLCO1B2 gene sequences described herein include but are not limited to the SLCO1B2 gene sequence from rat (genebank Gene ID: 58978), the SLCO1B2 gene sequence from mouse (genebank Gene ID: 28253), and the SLCO1B2 gene sequences from the above-mentioned other animals, including but not limited to gene bank NC_000072, NC_051339 and other SLCO1B2 gene sequences.

[0107] The invention also encompasses variants and homologous sequences of the aforementioned OATP1B2 amino acid sequence, nucleic acid coding sequence, and SLCO1B2 gene sequence. Herein, the agents or substances used to prevent or reduce the interaction between hepadnavirus and / or HDV and OATPs retain at least the activity of binding to or acting on OATPs, or acting on the region where HBV or HDV interacts with OATPs, thereby preventing or reducing infection by HBV and / or HDV. Such agents or substances with the activity of binding to or acting on OATPs include, but are not limited to, polypeptides derived from hepadnavirus surface antigens, polypeptides that bind to or act on OATPs, antibodies, proteins, small molecule drugs, nucleotides, and other agents, as well as currently known agents that act on OATPs. The methods for preventing or reducing the interaction between hepadnavirus and / or HDV and OATPs comprise administering to a subject in need thereof the agents or substances, including, but not limited to, polypeptides derived from hepadnavirus surface antigens, polypeptides that bind to or act on OATPs, antibodies, proteins, small molecule drugs, nucleotides, and other agents, as well as various currently known methods of acting on OATPs.

[0108] The agents and substances described herein can bind to or interact with OATPs, preventing or reducing the interaction between HBV and / or HDV and OATPs. The present invention specifically includes agents and substances that bind to or interact with OATP1B1 and / or OATP1B3, including but not limited to polypeptides derived from hepadnavirus surface antigens, polypeptides, antibodies, proteins, small molecule drugs, nucleotides, and other agents that can bind to or interact with OATP1B1 and / or OATP1B3, as well as currently known agents or substances that act on OATP1B1 and / or OATP1B3. These agents or substances can bind to or interact with OATP1B1 and / or OATP1B3, preventing or reducing the interaction between HBV and / or HDV and OATP1B1 and / or OATP1B3. The present invention specifically includes, but is not limited to, agents or substances that bind to or interact with human OATP1B1 and / or human OATP1B3, including, but not limited to, polypeptides derived from the surface antigen of hepadnavirus, polypeptides, antibodies, proteins, small molecule drugs, nucleotides, and other agents that can bind to or interact with human OATP1B1 and / or human OATP1B3, as well as currently known agents that act on human OATP1B1 and / or human OATP1B3. These agents can bind to or interact with human OATP1B1 and / or human OATP1B3 to prevent or reduce the interaction between HBV and / or HDV and human OATP1B1 and / or human OATP1B3.

[0109] The polypeptides described herein that can bind to or interact with OATPs include, but are not limited to, polypeptides or polypeptide modifications obtained by targeting OATPs using, but are not limited to, random peptide libraries, combinatorial peptide libraries, synthetic peptide libraries, and other peptide library technologies, phage display technology, yeast display technology, viral display technology, and other peptide screening technologies. In particular, they include, but are not limited to, polypeptides or polypeptide modifications that can bind to or interact with OATP1B1 and / or OATP1B3 that can be obtained by targeting OATP1B1 and / or OATP1B3 using, but are not limited to, random peptide libraries, combinatorial peptide libraries, synthetic peptide libraries, and other peptide library technologies, phage display technology, yeast display technology, viral display technology, and other peptide screening technologies. In particular, they include, but are not limited to, polypeptides or polypeptide modifications that can bind to or interact with human OATP1B1 and / or human OATP1B3 that can be obtained by targeting human OATP1B1 and / or human OATP1B3 using, but are not limited to, random peptide libraries, combinatorial peptide libraries, synthetic peptide libraries, and other peptide library technologies, phage display technology, yeast display technology, viral display technology, and other peptide screening technologies.

[0110] Antibodies capable of binding to or interacting with OATPs described herein include, but are not limited to, anti-OATP antibody serum, polyclonal antibodies, monoclonal antibodies, antibody fragments, or antibody modifications obtained by immunizing animals with, but not limited to, OATP proteins, protein fragments, or fusion proteins, or polyclonal antibodies, monoclonal antibodies, humanized antibodies, fully humanized antibodies, antibody fragments, fusion antibodies, bispecific antibodies, multispecific antibodies, and other antibodies, or antibody fragments, or antibody modifications obtained by hybridoma technology, phage display technology, yeast display technology, immune library technology, ribosome display and mRNA display screening technology, Nanobody (Nb) screening methods, transgenic mouse fully human antibody screening technology, single-cell photoluminescence technology, flow cytometry, and other antibody screening technologies. The above-mentioned antibodies or antibody serum are characterized by being able to bind to OATPs and prevent or reduce the interaction between HBV and / or HDV and OATPs. Specifically encompassed herein are antibody sera, polyclonal antibodies, monoclonal antibodies, humanized antibodies, fully humanized antibodies, antibody fragments, fusion antibodies, bispecific antibodies, multispecific antibodies, and other antibodies, or antibody fragments, or antibody modifications, that can bind to or interact with OATP1B1 and / or OATP1B3, and are characterized by being able to bind to OATP1B1 and / or OATP1B3 and prevent or reduce the interaction between HBV and / or HDV and OATP1B1 and / or OATP1B3. Specifically encompassed herein are antibody sera, polyclonal antibodies, monoclonal antibodies, humanized antibodies, fully humanized antibodies, antibody fragments, fusion antibodies, bispecific antibodies, multispecific antibodies, and other antibodies, or antibody fragments, or antibody modifications, that can bind to or interact with human OATP1B1 and / or human OATP1B3, and are characterized by being able to bind to human OATP1B1 and / or human OATP1B3 and prevent or reduce the interaction between HBV and / or HDV and human OATP1B1 and / or human OATP1B3. The present invention also includes antibodies, antibody fragments or antibody modifications targeting the region where HBV or HDV interacts with OATP.

[0111] The proteins described herein that can bind to or interact with OATPs include, but are not limited to, proteins, amino acid fragments, or protein modifications obtained by targeting OATPs through phage display technology, yeast display technology, cell display technology, random libraries, and other protein-protein interaction screening technologies. The present invention particularly includes, but is not limited to, proteins that can bind to or interact with OATP1B1 and / or OATP1B3, including, but not limited to, proteins, amino acid fragments, or protein modifications obtained by targeting OATP1B1 and / or OATP1B3 through phage display technology, yeast display technology, cell display technology, random libraries, and other protein-protein interaction screening technologies. The present invention particularly includes, but is not limited to, proteins that can bind to or interact with human OATP1B1 and / or human OATP1B3, including, but not limited to, proteins, amino acid fragments, or protein modifications obtained by targeting human OATP1B1 and / or human OATP1B3 through phage display technology, yeast display technology, cell display technology, random libraries, and other protein-protein interaction screening technologies.

[0112] The nucleotides described herein that can bind to or interact with OATPs include, but are not limited to, polyDNAs, oligoDNAs, polyRNAs, oligoRNAs, deoxyribonucleic acid-protein complexes (DNPs), or nucleic acid modifications obtained by screening with OATPs as targets. Specifically, but are not limited to, nucleotides that can bind to or interact with OATP1B1 and / or OATP1B3, including, but not limited to, polyDNAs, oligoDNAs, polyRNAs, oligoRNAs, deoxyribonucleic acid-protein complexes (DNPs), or nucleic acid modifications obtained by screening with OATP1B1 and / or OATP1B3 as targets. Specifically, but are not limited to, nucleotides that can bind to or interact with human OATP1B1 and / or human OATP1B3, including, but not limited to, polyDNAs, oligoDNAs, polyRNAs, oligoRNAs, deoxyribonucleic acid-protein complexes (DNPs), or nucleic acid modifications obtained by screening with human OATP1B1 and / or human OATP1B3 as targets.

[0113] The small molecule compounds described herein that can bind to or interact with OATPs include, but are not limited to, small molecule compounds that bind to or interact with OATPs obtained by design or compound screening. Specifically, but are not limited to, small molecule compounds that can bind to or interact with OATP1B1 and / or OATP1B3, including, but are not limited to, small molecule compounds that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by design or compound screening. Specifically, but are not limited to, small molecule compounds that can bind to or interact with human OATP1B1 and / or human OATP1B3, including, but are not limited to, small molecule compounds that can bind to or interact with human OATP1B1 and / or human OATP1B3 obtained by design or compound screening.

[0114] The agents and substances known to act on OATP described herein include, but are not limited to, agents and substances known to act on OATP1B1 and / or OATP1B3, and particularly include, but are not limited to, agents and substances known to act on human OATP1B1 and / or human OATP1B3, such agents and substances include, but are not limited to, statins (including, but not limited to, atorvastatin, pitavastatin, rosuvastatin, simvastatin, pravastatin and other statins), sartans (including, but not limited to, valsartan, olmesartan, telmisartan and other sartans), bosentan, docetaxel, fexofenadine, glecaprevir, glyburide (glibenclamide), grazoprevir, nateglinide, paclitaxel, paritaprevir, repaglinide, simeprevir, voxilaprevir, steroid hormone metabolites, thyroid hormones metabolites, inflammatory mediators, bile acids, bilirubin, bromosulfophthalein, estradiol-17β-glucuronide, cholecystokinin octapeptide, estradiol-3-sulfate, liver function markers, mushroom toxins, antibiotics, antiviral drugs, anticancer drugs, and other agents and substances known to interact with OATP1B1 and / or OATP1B3.

[0115] In the prevention or reduction of OATP expression and / or function described herein, the agents or substances used at least retain their effect on the OATP encoding gene SLCO, or on OATP mRNA, or on OATP protein expression, and / or modification, and / or transport, and / or degradation, to prevent or reduce the expression level of OATP protein, or reduce the biological function of the expressed OATP protein, or enhance OATP protein degradation. The inhibition or reduction of OATP expression and / or function described herein specifically includes, but is not limited to, inhibition or reduction of OATP1B1 and / or OATP1B3 expression and / or function, at least retaining the effect on the OATP1B1 and / or OATP1B3 encoding genes SLCO1B3 or SLCO1B1, or on OATP1B1 and / or OATP1B3 mRNA, or on OATP1B1 and / or OATP1B3 protein expression, and / or modification, and / or transport, and / or degradation, to inhibit or reduce the expression level of OATP1B1 and / or OATP1B3 proteins, or reduce the biological function of the expressed OATP1B1 and / or OATP1B3 proteins, or enhance the degradation of OATP1B1 and / or OATP1B3 proteins. The inhibition or reduction of OATP1B1 and / or OATP1B3 expression and / or function described herein specifically includes, but is not limited to, inhibition or reduction of human OATP1B1 and / or human OATP1B3 expression and / or function, while at least retaining an effect on the human OATP1B1 and / or human OATP1B3 encoding genes, human SLCO1B3 or human SLCO1B1, or on human OATP1B1 and / or human OATP1B3 mRNA, or on human OATP1B1 and / or human OATP1B3 protein expression, and / or modification, and / or transport, and / or degradation, to inhibit or reduce the expression level of human OATP1B1 and / or human OATP1B3 proteins, or reduce the biological functions of the expressed human OATP1B1 and / or human OATP1B3 proteins, or enhance the degradation of human OATP1B1 and / or human OATP1B3 proteins.

[0116] The methods described herein for acting on the SLCO gene include, but are not limited to, gene knockout, gene insertion, homologous recombination, gene editing, and other methods for inhibiting gene transcriptional activity. The methods described herein for acting on the SLCO gene include, but are not limited to, methods for acting on the SLCO1B3 or SLCO1B1 gene, and in particular, but are not limited to, methods for acting on the human SLCO1B3 or human SLCO1B1 gene, including, but not limited to, gene knockout, gene insertion, homologous recombination, gene editing, and other methods for inhibiting gene transcriptional activity. The methods described herein for acting on the OATP mRNA include, but are not limited to, antisense nucleic acids, RNA interference, and other methods for cutting and degrading mRNA, destroying mRNA stability, or preventing mRNA translation. The methods described herein for acting on the OATP mRNA include, but are not limited to, methods for acting on OATP1B1 and / or OATP1B3 mRNA, and in particular, but are not limited to, methods for acting on human OATP1B1 and / or human OATP1B3 mRNA, including, but not limited to, antisense nucleic acids, RNA interference, and other methods for cutting and degrading mRNA, destroying mRNA stability, or preventing mRNA translation.

[0117] The methods described herein for reducing the biological function of the expressed OATP protein include, but are not limited to, inhibiting post-translational modifications such as protein glycosylation to reduce the biological function of the OATP protein. The methods for reducing the biological function of the expressed OATP protein specifically include, but are not limited to, methods for reducing the biological function of the expressed OATP1B1 and / or OATP1B3 protein, and particularly include, but are not limited to, methods for reducing the biological function of the expressed human OATP1B1 and / or human OATP1B3 protein, including, but not limited to, inhibiting post-translational modifications such as protein glycosylation to reduce the biological function of the human OATP1B1 and / or human OATP1B3 protein. The methods for acting on the transport of OATP proteins include, but are not limited to, methods for inhibiting protein sorting and the Golgi apparatus to reduce the amount of OATP protein presented on the cell membrane surface, or methods for enhancing the endocytosis of OATP protein on the cell membrane surface to reduce the amount of OATP protein on the cell membrane surface.

[0118] The methods described herein for acting on OATP protein transport specifically include, but are not limited to, methods for acting on OATP1B1 and / or OATP1B3 protein transport, and particularly include, but are not limited to, methods for acting on human OATP1B1 and / or human OATP1B3 protein transport, including but not limited to methods for inhibiting protein sorting and the Golgi apparatus to reduce the amount of human OATP1B1 and / or human OATP1B3 protein presented on the cell membrane surface, or methods for enhancing the endocytosis of human OATP1B1 and / or human OATP1B3 protein on the cell membrane surface to reduce the amount of human OATP1B1 and / or human OATP1B3 protein on the cell membrane surface.

[0119] The methods for enhancing OATP protein degradation described herein particularly include, but are not limited to, methods for enhancing OATP1B1 and / or OATP1B3 protein degradation, and particularly include, but are not limited to, methods for enhancing human OATP1B1 and / or human OATP1B3 protein degradation, including but not limited to increasing protein ubiquitination, protein degradation targeting chimeras (PROTACs), and other methods for disrupting protein stability and / or enhancing protein degradation.

[0120] The methods, agents, or substances described herein for inhibiting or reducing the expression and / or function of OATP1B1 and / or OATP1B3 further include, but are not limited to, gene editing targeting the SLCO gene, antisense nucleic acids targeting OATP mRNA, ribozymes, DNAzymes, or RNA interference, and PROTACs targeting OATP proteins. Gene editing, also known as genome editing, includes but is not limited to using Cre-lox-mediated recombination at specific genomic sites, zinc finger nuclease-directed modification of target genes, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) / Cas9 systems and technology variants, or other gene editing technologies to edit (directed modification) the SLCO gene and its transcripts, thereby adding or deleting DNA fragments, deleting or replacing specific DNA bases, and thereby changing the sequence, expression level, or function of the SLCO gene or regulatory elements. In particular, the SLCO1B3 and / or SLCO1B1 genes and their transcripts are edited (directed modification) to alter the sequence, expression level, or function of the SLCO1B3 and / or SLCO1B1 genes or regulatory elements. In particular, the human SLCO1B3 and / or human SLCO1B1 genes and their transcripts are edited (directed modification) to alter the sequence, expression level, or function of the human SLCO1B3 and / or human SLCO1B1 genes or regulatory elements. RNA interference (RNAi) includes, but is not limited to, methods and agents for degrading OATP mRNA using double-stranded RNA (dsRNA), small interfering RNA (siRNA), silencing RNA, or RNA interference vectors, and particularly includes, but is not limited to, methods and agents for degrading OATP1B1 and / or OATP1B3 mRNA, and particularly includes, but is not limited to, methods and agents for degrading human OATP1B1 and / or human OATP1B3 mRNA. The methods and agents for PROTAC targeting OATP proteins include but are not limited to methods and agents for degrading the target protein OATP using the ubiquitin-proteasome system (UPS), and the methods and agents may contain a ubiquitin ligase ligand-linker-target protein OATP ligand structure, or other structures and methods that can ubiquitinate or direct OATP to the proteasome.The methods and agents for PROTAC targeting OATP proteins particularly include but are not limited to methods and agents for PROTAC targeting OATP1B1 and / or OATP1B3 proteins, and particularly include but are not limited to methods and agents for PROTAC targeting human OATP1B1 and / or human OATP1B3 proteins.

[0121] The methods, agents, and substances described herein for treating and / or preventing HBV and / or HDV infection and related diseases by preventing or reducing the interaction between HBV and / or HDV and OATP or by preventing or reducing the expression and / or function of OATP can be used alone or in combination with agents that prevent or reduce the interaction between HBV and / or HDV and NTCP and / or agents and methods that can prevent or reduce the expression and / or function of NTCP in animals.

[0122] The NTCPs described herein include, but are not limited to, mammals, birds, fish, amphibians, and other animals. Mammals include, but are not limited to, primates, tree shrews, rodents, bats, even-toed ungulates, carnivores, and other mammals. Birds include, but are not limited to, ducks, herons, and other avian animals. Primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include but are not limited to crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include but are not limited to Sciuridae animals, myomorphs and other rodents. The Sciuridae animals include but are not limited to arctic ground squirrels, ground squirrels, woodchucks and other Sciuridae animals. The myomorphs include but are not limited to rats (Rattus rattus), mice (Mus musculus) and other myomorphs. The Artiodactyla animals include but are not limited to pigs and other even-toed ungulates. The Carnivora animals include but are not limited to dogs and other Carnivora animals.In some embodiments of the present invention, the NTCP includes but is not limited to NTCP from humans (SEQ ID NO: 7, NCBI Reference Sequence: NM_003049), NTCP from chimpanzees (NCBI Reference Sequence: XM_510035), NTCP from tree shrews (NCBI Reference Sequence: XM_006171503), NTCP from bonobos (NCBI Reference Sequence: XM_003824101), NTCP from capuchin monkeys (NCBI Reference Sequence: XM_032245781), NTCP from squirrel monkeys (NCBI Reference Sequence: XM_003924480), NTCP from cynomolgus monkeys (NCBI Reference Sequence: NM_001283323), NTCP from arctic ground squirrels (NCBI Reference Sequence: XM_026385176), NTCP from ground hamsters (NCBI Reference Sequence: XM_026385177), and NTCP from ground hamsters (NCBI Reference Sequence: XM_003824101). Sequence: XM_005322843), NTCP from rat (NCBI Reference Sequence: XM_032908052), NTCP from mouse (NCBI Reference Sequence: NM_001177561), NTCP from porcine (NCBI Reference Sequence: XM_001927695), NTCP from dog (NCBI Reference Sequence: XM_537494), as well as NTCP and variants thereof from the other animals mentioned above.

[0123] In some preferred embodiments of the methods, agents, and substances herein for preventing or reducing the interaction between HBV and / or HDV and OATP, or for treating and / or preventing HBV and / or HDV infection and related diseases by preventing or reducing OATP expression and / or function, when used in combination with an agent that prevents or reduces the interaction between HBV and / or HDV and NTCP and / or an agent and method that can prevent or reduce NTCP expression and / or function in animals, the NTCP is human, primate, rodent, or avian NTCP, more preferably human NTCP. The agents and substances that prevent or reduce the interaction between HBV and / or HDV and NTCP and / or the agents, substances, and methods that prevent or reduce NTCP expression and / or function in animals include, but are not limited to, polypeptides, proteins, antibodies, small molecule drugs, polynucleotides, and other agents that interact with NTCP or HBV to thereby prevent or reduce the interaction between HBV and / or HDV and NTCP, as well as currently known agents, substances, and methods that act on NTCP. The currently known agents, substances and methods for acting on NTCP include but are not limited to bile acids and their derivatives (including but not limited to bile acid CA, deoxycholic acid DCA, chenodeoxycholic acid CDCA, ursodeoxycholic acid UDCA, lithocholic acid LCA and products of these bile acids combined with glycine, taurine, sulfuric acid, glucuronic acid, dimeric bile acid derivatives), organic anions (including but not limited to glycocholic acide, bromosulfophthalein), polypeptides (including but not limited to EMD51921, angiopeptin, U71038, α-amanitln, phe-phe-phe, ala-ala-ala, tyr-tyr), drugs (including but not limited to propranolol, progesterone, cyclosporine and its derivatives, ketoconazole, bumetanide, furosemide, chlorpropamide, theophylline, ephedrine, sodium valproate, nicotine, pseudoephedrine, salicylate, procainamide, diphenylhydantoin, nifedipine, caffeine, bupivacaine, lidocaine, quinidlne and other drugs that interact with NTCP) and other agents and methods that interact with NTCP.

[0124] The agents, substances, and methods described herein for preventing or reducing NTCP expression and / or function at least retain an effect on the NTCP encoding gene SLC10A1, or on NTCP mRNA, or on NTCP protein expression, and / or modification, and / or transport, and / or degradation, to prevent or reduce NTCP protein expression, or reduce the biological function of the expressed NTCP protein, or enhance NTCP protein degradation. The methods for acting on the SLC10A1 gene include, but are not limited to, gene knockout, gene insertion, homologous recombination, gene editing, and other methods for inhibiting gene transcriptional activity. The methods for acting on NTCP mRNA include, but are not limited to, antisense nucleic acids, RNA interference, and other methods for cutting and degrading mRNA, disrupting mRNA stability, or preventing mRNA translation. The methods for reducing the biological function of the expressed NTCP protein include, but are not limited to, inhibiting post-translational modifications such as protein glycosylation to reduce the biological function of the NTCP protein. The methods for acting on NTCP protein transport include, but are not limited to, methods such as inhibiting protein sorting and the Golgi apparatus to reduce the amount of NTCP protein presented on the cell membrane surface, or methods such as enhancing the endocytosis of NTCP protein on the cell membrane surface to reduce the amount of NTCP protein on the cell membrane surface. The method for enhancing NTCP protein degradation includes but is not limited to increasing protein ubiquitination, protein degradation targeting chimera (PROteolysis TArgeting Chimera, PROTAC) and other methods of destroying protein stability and / or enhancing protein degradation.

[0125] The methods, agents, and substances described herein for preventing or reducing the interaction of HBV and / or HDV with OATPs, or by preventing or reducing the expression and / or function of OATPs, can be used to prevent and treat HBV and / or HDV infection and related diseases. In some embodiments, the disease is a liver disease associated with HBV and / or HDV viral infection. The various methods, agents, and substances described herein can be used to block the transport of HBV and / or HDV into cells via OATPs, thereby preventing or reducing viral infection of liver cells and blocking the onset and / or progression of the disease. In the event of accidental viral exposure, the methods, agents, and substances described herein can be administered to block the entry of HBV and / or HDV into liver cells, thereby blocking viral infection and preventing the disease. Therefore, the methods for treating or preventing liver disease described herein are methods for treating or preventing liver disease caused by viral infection, comprising administering to a subject in need thereof a therapeutically or prophylactically effective amount of the various agents or substances described herein for preventing or reducing the interaction of HBV and / or HDV with OATPs, or by preventing or reducing the expression and / or function of OATPs. In other embodiments, provided herein is a method for blocking HBV and / or HDV virus infection of liver tissue or liver cells and / or blocking the spread of the virus between liver cells, the method comprising administering to a subject in need thereof an effective amount of the various agents or substances described herein that prevent or reduce the interaction of HBV and / or HDV with OATP, or by preventing or reducing the expression and / or function of OATP. In some embodiments, provided herein are uses of the various substances or agents described herein in the preparation of a medicament for treating or preventing liver disease, especially a liver disease associated with hepatitis B virus or hepatitis D virus infection, and in the preparation of a medicament for blocking a virus (such as a hepatitis virus, such as hepatitis B virus, hepatitis D virus) from infecting liver tissue or liver cells and / or blocking the spread of the virus between liver cells. In some embodiments, provided herein are methods, agents and substances for treating or preventing liver disease, especially a liver disease associated with hepatitis B virus or hepatitis D virus infection, as well as methods, agents and substances for blocking a virus (such as a hepatitis virus, such as hepatitis B virus or hepatitis D virus) from infecting liver tissue or liver cells and / or blocking the spread of the virus between liver cells. In some embodiments, the disease is acute hepatitis B (AHB), chronic hepatitis B (CHB), acute hepatitis D (AHD), or chronic hepatitis D (CHD).

[0126] In some embodiments, the methods, agents, or substances for treating or preventing HBV and / or HDV infection and related diseases described herein can be used in combination with other HBV and / or HDV treatment or prevention drugs, including but not limited to nucleoside (acid) analogs (including but not limited to lamivudine, telbivudine, entecavir, tenofovir disoproxil, adefovir dipivoxil, clavudine, tenofovir disoproxil and other nucleoside (acid) analogs), interferons (including but not limited to conventional interferons (including but not limited to interferon α-2a, α-2b and other types of interferons), long-acting interferons, PEG interferons (including but not limited to PEG interferon α-2a, α-2b and other types of PEG interferons) and other interferons), nucleocapsid inhibitors, RNA interference, immunomodulators and other agents for preventing or treating HBV or HDV infection, as well as combined use regimens of these agents.

[0127] In some embodiments of the present invention, the agent or substance that prevents or reduces the interaction between hepadnavirus and / or HDV and OATP is selected from a peptide derived from the surface antigen of hepadnavirus that binds to OATP. Preferably, the peptide derived from the surface antigen of hepadnavirus that binds to OATP is as described below.

[0128] OATP-binding hepadnavirus surface antigen-derived peptide

[0129] Also provided herein are polypeptides derived from a hepadnavirus and variants thereof, which retain at least the biological activity of binding to an OATP and are referred to herein as OATP-binding peptides. "Derived from a hepadnavirus," as used herein, means that the polypeptide is derived from a hepadnavirus, or is highly homologous to or has the same function as a corresponding polypeptide from a hepadnavirus but has certain sequence differences. "Variants," as used herein, refer to polypeptides that have certain amino acid differences compared to a wild-type polypeptide but still possess the biological activity of binding to an OATP as described herein. The OATP-binding peptides described herein can be used to prevent and / or treat hepadnavirus and / or HDV infection and related diseases.

[0130] The hepadnaviruses described herein include, but are not limited to, orthohepadnavirus, avihepadnavirus, metahepadnavirus, herpetohepadnavirus, and other hepadnaviruses. The orthohepadnaviruses include, but are not limited to, orthohepadnaviruses that infect humans (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkeys, woodchuck hepatitis virus (WHV), Arctic ground shrews, ground shrews (Ground squirrel hepatitis virus, GSHV), tree shrews, chimpanzees, bats (BHV), and other animals. Exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers AF046996 (WMHBV), NC_043528 (CMHBV), NC_004107 (WHV), NC_001484 (GSHV), and JX941468 (BHV). The Avihepadnavirus includes but is not limited to Avihepadnavirus that infects ducks (duck HBV, DHBV), herons (heron HBV, HHBV) and other birds. Exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers NC_001344 (DHBV) and NC_001486 (HHBV).

[0131] The original hepadnavirus (HBV) that infects humans includes, but is not limited to, genotypes A, B, C, D, E, F, G, H, and I HBV. Exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G), and AB179747 (genotype H). In some embodiments herein, the amino acid sequence of the hepadnavirus-derived peptide that binds to OATP is derived from a viral envelope protein, also known as a surface antigen or surface protein, i.e., a hepadnavirus surface antigen-derived peptide that binds to OATP or an OATP-binding peptide derived from a hepadnavirus surface antigen.

[0132] In a further preferred embodiment herein, the amino acid sequence of the hepadnavirus surface antigen-derived peptide that binds to OATP is derived from the amino acid sequence of the viral surface antigen Pre-S region, including but not limited to the exemplary surface antigen Pre-S amino acid sequences SEQ ID NOs: 8-22 and variants thereof. Specifically encompassed herein are polypeptides derived from the amino acid sequence of the human HBV surface antigen Pre-S 1 region (i.e., HBVPre-S1-derived peptides that bind to OATP or OATP-binding peptides derived from HBVPre-S1), including but not limited to the amino acid sequences of the HBVPre-S1 region of genotypes A, B, C, D, E, F, G, H, and I, including but not limited to the exemplary HBV surface antigen Pre-S amino acid sequences SEQ ID NOs: 15-22 and variants thereof. Specifically encompassed herein are OATP-binding peptides derived from the amino acid sequence of the human HBV surface antigen Pre-S1 region, including but not limited to the amino acid sequence of the HBVPre-S region of genotype C, including but not limited to the exemplary HBV surface antigen Pre-S1 region amino acid sequence SEQ ID NO: 17 and variants thereof. Hepadnavirus-derived peptides and variants thereof that bind to OATP, hepadnavirus surface antigen-derived peptides and variants thereof that bind to OATP, and HBVPre-S1-derived peptides and variants thereof that bind to OATP are collectively referred to herein as OATP-binding peptides.

[0133] The hepadnavirus-derived peptides and variants thereof that bind to OATPs at least retain biological activity in binding to OATPs. The OATPs include, but are not limited to, the OATP superfamily from mammals, birds, fish, amphibians, and other animals, including, but not limited to, the OATP1, OATP2, OATP3, OATP4, OATP5, OATP6, OATP7, OATP8, OATP9, OATP10, and OATP11 families, particularly including, but not limited to, the OATP family expressed in hepatocytes; further including, but not limited to, the OATP1B subfamily, particularly including, but not limited to, the OATP1B subfamily expressed in hepatocytes; and further including, but not limited to, OATP1B3, OATP1B1, OATP1B2, and other OATP1B subfamily members. The OATP1B3 includes, but is not limited to, OATP1B3 from humans (NCBI Reference Sequence: NM_019844), OATP1B3 from chimpanzees (NCBI Reference Sequence: XM_016923821), OATP1B3 from tree shrews (NCBI Reference Sequence: XM_006155085), OATP1B3 from cynomolgus monkeys (NCBI Reference Sequence: NM_001283191), OATP1B3 from Arctic ground squirrels (NCBI Reference Sequence: XM_026391932), OATP1B3 from rats (NCBI Reference Sequence: XM_032905480), OATP1B3 from pigs (NCBI Reference Sequence: NM_001315607), and OATP1B3 from dogs (NCBI Reference Sequence: XM_001315608). Sequence: NM_001166047), and OATP1B3 from other animals. The OATP1B1 includes but is not limited to OATP1B1 from humans (NCBI Reference Sequence: NM_006446), OATP1B1 from bonobos (NCBI Reference Sequence: XM_003828901), OATP1B1 from cynomolgus monkeys (NCBI Reference Sequence: NM_001284540), and OATP1B1 from other animals.The OATP1B2 includes, but is not limited to, rat OATP1B2 (NCBI Reference Sequence: NM_031650), mouse OATP1B2 (NCBI Reference Sequence: NM_020495), and other animal OATP1B2. In some preferred embodiments herein, the OATP-binding HBV Pre-S1-derived peptide and variants thereof at least retain the biological activity of binding to human OATP1B3 and / or human OATP1B1.

[0134] The term "hepadnavirus-derived peptide that binds to OATP" or "OATP-binding peptide derived from hepadnavirus" means that the origin or source of the polypeptide is a hepadnavirus, including but not limited to natural, recombinant, synthetic or purified polypeptides, including but not limited to full-length natural HBV polypeptides or fragments thereof, and variants of full-length natural polypeptides or fragments thereof. In some embodiments, the hepadnavirus-derived peptide that binds to OATP can be derived from any hepadnavirus, including but not limited to orthohepadnavirus (OHV) that infects mammals, avihepadnavirus (AHV) that infects birds, Metahepadnavirus that infects fish, and Herpetohepadnavirus that infects amphibians; in some embodiments, the hepadnavirus surface antigen-derived peptide that binds to OATP can be derived from any hepadnavirus surface antigen; in some embodiments, the hepadnavirus surface antigen-derived peptide that binds to OATP can be derived from any hepadnavirus surface antigen; The hepadnavirus surface antigen-derived peptide that binds to OATP can be derived from any original hepadnavirus surface antigen; in some embodiments, the hepadnavirus surface antigen-derived peptide that binds to OATP can be derived from any human hepatitis B virus (HBV) surface antigen, including but not limited to HBV surface antigens derived from genotypes A, B, C, D, E, F, G, H and I; in some embodiments, the hepadnavirus surface antigen-derived peptide that binds to OATP can be derived from any human hepatitis B virus (HBV) surface antigen Pre-S1 region, an HBVPre-S1-derived peptide that binds to OATP, or an OATP-binding peptide derived from HBVPre-S1.

[0135] The terms "HBVPre-S1-derived peptide that binds to OATP" or "OATP-binding peptide derived from HBVPre-S1" refer to polypeptides derived from the HBV surface antigen Pre-S1 region, including but not limited to natural, recombinant, synthetic, or purified polypeptides, including but not limited to the full-length natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, full-length non-natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, non-full-length natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, non-full-length non-natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, and other forms of variants. In some embodiments, the OATP-binding HBVPre-S1-derived peptide may comprise the entire surface antigen pre-S1 region of any HBV genotype, starting from amino acid G (glycine) at position 2 of the surface antigen pre-S1 region. In certain embodiments, the OATP-binding HBVPre-S1-derived peptides described herein can be derived from the pre-S1 region of the surface antigen of any one of HBV genotypes A, B, C, D, E, F, G, H, and I. Exemplary genomic sequences of these HBV genotypes can be found, but are not limited to, GenBank accession numbers KC875260, AY220704, AF461363, AY796030, AB205129, DQ823095, HE981176, and AB179747, and exemplary amino acid sequences of the pre-S1 regions of their surface antigens can be found, but are not limited to, SEQ ID NOs: 8-22 and variants thereof. In certain embodiments, the OATP-binding HBVPre-S1-derived peptides described herein can be derived from the pre-S1 region of the surface antigen of HBV genotype C. The OATP-binding HBVPre-S1 derivative peptide at least retains the biological activity of binding to OATP, particularly retains the biological activity of binding to OATP1B1, and / or OATP1B2 and / or OATP1B3, and especially retains the biological activity of binding to human OATP1B1 and / or human OATP1B3.

[0136] In some embodiments, the OATP-binding peptides described herein may be 10-118 amino acids in length. For example, the length of the polypeptide may be, but is not limited to, 15-100, 15-80, 20-100, 20-80, 20-60, 25-60, 30-60, 35-60, or 40-60 (including all integers between these ranges) amino acids. In some embodiments, the length of the OATP-binding peptides described herein may be at least 20 amino acids, such as at least 25, 30, 35, or 40 amino acids. In some embodiments, the length of the OATP-binding peptides described herein may be, but is not limited to, 20, 25, 30, 35, 40, 47, 55, or 60 amino acids. In some embodiments, the length of the OATP-binding peptides described herein may be 47 amino acids. The variants of the OATP-binding peptides described herein, which have different lengths, retain one or more biological activities associated with the corresponding polypeptides, including at least the biological activity of binding to OATPs, particularly the biological activity of binding to OATP1B1, and / or OATP1B2 and / or OATP1B3, and especially the biological activity of binding to human OATP1B1 and / or human OATP1B3.

[0137] In certain embodiments, the OATP-binding peptides described herein may be derived from the pre-S1 region of the genotype C HBV surface antigen, including but not limited to amino acids 2-119, 2-69, 2-59, 13-119, 13-88, 13-72, 13-67, 13-59, 13-52, 13-47, 13-42, 13-37, 13-32 of the pre-S1 region and variants thereof, exemplary sequences of which include but are not limited to SEQ ID NOs: 23-35 and variants thereof.

[0138] In some embodiments, the OATP-binding peptide comprises the amino acid sequence of at least amino acid residues 13-44 at the N-terminus of the pre-S1 region of the genotype C HBV surface antigen as described herein, and variants thereof. In preferred embodiments, the OATP-binding peptide may be 32-47 amino acid residues in length. In particularly preferred embodiments, the exemplary amino acid sequence of the OATP-binding peptide is as shown in SEQ ID NO: 30; preferably, the OATP-binding peptide also includes variants of the amino acid sequence shown in SEQ ID NO: 30; in some embodiments, the OATP-binding peptide comprises at least amino acid residues 1-32 of the exemplary sequence SEQ ID NO: 30, and variants thereof, preferably, the OATP-binding peptide comprises at least amino acid residues 1 to 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, or 46 of the exemplary sequence SEQ ID NO: 30, and variants thereof. In some embodiments, the OATP-binding peptide refers to a fragment containing at least amino acid residues 13-44 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I, and variants thereof, a fragment containing at least amino acid residues 2-33 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and variants thereof, or a fragment containing at least amino acid residues 12-43 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E and G, and variants thereof. In some embodiments, the OATP-binding peptide comprises a fragment containing at least amino acid residues 13-69 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I, and variants thereof, a fragment containing at least amino acid residues 2-48 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and variants thereof, and a fragment containing at least amino acid residues 12-68 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E or G, and variants thereof, exemplary sequences thereof include, but are not limited to, exemplary sequences of SEQ ID NOs: 36-42 and variants thereof.

[0139] As used herein, a "variant" with respect to an OATP-binding peptide refers to a peptide that differs from a given polypeptide in amino acid sequence but retains biological activity for binding to an OATP, particularly biological activity for binding to OATP1B3, and / or OATP1B2, and / or OATP1B1, and particularly biological activity for binding to human OATP1B3 and / or OATP1B1. For example, in some embodiments, one or a combination of two or more of the following amino acid substitutions can be introduced into a derivative peptide derived from the amino acid sequence of positions 13-59 of the HBVPre-S1 region of genotype C: N15D, F25L, G35K, N39E, F45L, N46K, N48H or N48Y or N48K, D50A, H51Q or H51N, E54K or E54D, A55S, N56K or N56D, and Q57K. Exemplary sequences include, but are not limited to, those set forth in SEQ ID NOs: 43-60. The term "variant" also includes homologous polypeptide sequences found in different viral species, strains, or subtypes of the genus Hepadnavirus. HBV is divided into four major serotypes (adr, adw, ayr, and ayw) based on antigenic epitopes on its envelope protein, and into nine genotypes (AI) based on the variability of the entire nucleotide sequence in the genome. Therefore, the term "variant" includes any of these homologous polypeptides found in HBV subtypes. "Variant" also includes polypeptides and variants thereof with naturally occurring flanking amino acid sequences from any of these HBV subtypes added to the N- and / or C-termini. For example, in some embodiments, a polypeptide derived from amino acid sequences 13-59 of the pre-S1 region of the genotype C surface antigen can be fused with, but not limited to, a fragment comprising at least amino acid residues 2-12 of the N-terminal pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, or I, or a fragment comprising at least amino acid residues 2-11 of the N-terminal pre-S1 region of the HBV surface antigen of genotypes E and G to form a fusion polypeptide. Example sequences include, but are not limited to, those set forth in SEQ ID NOs: 61-66. For example, in some embodiments, the OATP-binding peptides described herein may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66, and a naturally occurring amino acid sequence flanking the N- and / or C-terminus of a surface antigen of any one of HBV genotypes A-I. In some embodiments, the naturally occurring flanking amino acid sequence may be derived from, but is not limited to, an amino acid sequence of an HBV surface antigen having GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G), or AB179747 (genotype H).For example, the polypeptide described herein may contain the amino acid sequence set forth in SEQ ID NO: 30 and contain, at its N- and / or C-terminus, a naturally occurring flanking amino acid sequence from the pre-S1 region of HBV genotype C. Alternatively, the OATP-binding peptide described herein may contain the amino acid sequence set forth in SEQ ID NO: 30 and contain, at its N- and / or C-terminus, a naturally occurring flanking amino acid sequence from the pre-S1 region of any one of HBV genotypes A, B, C, D, E, F, G, and H. In some embodiments, the N- and / or C-terminus of the polypeptide described herein may independently contain a naturally occurring flanking amino acid sequence of 1-10 amino acids, such as 1-8, 1-5, or 1-3 amino acids in length (including all integers within these ranges). For example, the OATP-binding peptide described herein may contain the amino acid sequence set forth in SEQ ID NO: 30 and contain, at its N-terminus, a naturally occurring flanking amino acid sequence of 10 amino acids from the pre-S1 region of HBV genotype C. In other words, the OATP-binding peptide may comprise amino acids 2-59 of the pre-S1 region of HBV genotype C (SEQ ID NO: 25). As another example, the OATP-binding peptide described herein may comprise the amino acid sequence set forth in SEQ ID NO: 30 and, at its N-terminus, a naturally occurring flanking amino acid sequence of 9 amino acids from the pre-S1 region of HBV genotype E. In other words, the OATP-binding peptide may comprise amino acids 13-59 of the pre-S1 region of HBV genotype C and amino acids 2-11 of the pre-S1 region of HBV genotype E (SEQ ID NO: 63). It should be understood that any of the OATP-binding peptides described herein may have naturally occurring flanking amino acid sequences of any length extending from its N- and / or C-terminus, and the resulting polypeptide retains the biological activity of the original polypeptide in binding to OATPs, particularly in binding to OATP1B3, and / or OATP1B2, and / or OATP1B1, and particularly in binding to human OATP1B3 and / or OATP1B1. "Variants" of the OATP-binding peptides also include those having one or more amino acid deletions, substitutions, or insertions while retaining biological activity in binding to OATPs, particularly biological activity in binding to OATP1B3, and / or OATP1B2, and / or OATP1B1, and particularly biological activity in binding to human OATP1B3 and / or OATP1B1. The OATP-binding peptide preferably retains the glycine corresponding to amino acid position 13 in the pre-S1 region of genotype C HBV (i.e., the N-terminal glycine of SEQ ID NO: 30). In some embodiments, the polypeptides described herein may have one or more naturally occurring mutations in the pre-S1 region of HBV.For example, in some embodiments, the OATP-binding peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the sequence from the pre-S1 region of HBV. In some embodiments, the OATP-binding peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to any one of the exemplary amino acid sequences selected from SEQ ID NOs: 23-66. In some embodiments, the OATP-binding peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the exemplary amino acid sequence of SEQ ID NO: 30. In some embodiments, the OATP-binding peptides described herein may have 1-3 amino acid deletions, substitutions, or insertions relative to the exemplary amino acid sequence of SEQ ID NO: 30. In certain embodiments, the OATP-binding peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions or insertions at the C-terminus relative to any of the exemplary amino acid sequences selected from SEQ ID NOs: 23-66. For example, the OATP-binding peptides may include, but are not limited to, an amino acid sequence selected from any of the exemplary amino acid sequences of SEQ ID NOs: 31-35. In various embodiments, the present invention includes, but is not limited to, the polypeptides described herein, as well as polypeptides having at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any of the polypeptides described. For example, the OATP-binding peptide may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to any of the exemplary sequences in SEQ ID NOs: 23-66; preferably, the amino acid sequence having such identity is derived from any of the hepadnaviruses described herein, particularly from a protohepadnavirus, more preferably from a protohepadnavirus (HBV) that infects humans, including but not limited to genotypes A, B, C, D, E, F, G, H and I HBV, more preferably from genotype C.In some embodiments, the OATP-binding peptide may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence exemplified in SEQ ID NO: 30; preferably, the amino acid sequence having such identity is derived from any of the hepadnaviruses described herein, particularly from a protohepadnavirus, more preferably from a protohepadnavirus (HBV) that infects humans, including but not limited to genotypes A, B, C, D, E, F, G, H, and I HBV, more preferably from genotype C. Variants having a certain sequence identity to the OATP-binding peptides described herein retain one or more biological activities of the corresponding polypeptide, including at least the biological activity of binding to OATPs, particularly the biological activity of binding to OATP1B3, and / or OATP1B2, and / or OATP1B1, and particularly the biological activity of binding to human OATP1B3 and / or OATP1B1.

[0140] The OATP-binding hepadnavirus-derived peptides and variants thereof are modified at their N-termini with a hydrophobic group, such as, but not limited to, myristic acid (myr), palmitic acid (plam), stearic acid (stearoyl), cholesterol (chol), oleic acid, linoleic acid, polyethylene glycol (PEG), arachidonic acid, or other hydrophobic groups. In some embodiments, the hydrophobic group can be selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In some embodiments, the hydrophobic group can be myristic acid. In certain embodiments, the OATP-binding peptides described herein may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, wherein the N-terminus of the OATP-binding peptide may be modified with a hydrophobic group selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In certain embodiments, the polypeptides may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, wherein the N-terminus of the OATP-binding peptide may be myristoylated. In some embodiments, the OATP-binding peptide described herein may comprise the amino acid sequence set forth in SEQ ID NO: 30 or SEQ ID NO: 60, wherein the N-terminus of the OATP-binding peptide may be myristoylated. In some embodiments, the C-terminus of the polypeptide may be modified. The C-terminal modification may be selected from, but not limited to, amidation (amination), prenylation, and other C-terminal modifications, or may be absent. In some embodiments, the C-terminal modification may be amidation (NH2). For example, the OATP-binding peptide may comprise the amino acid sequence set forth in SEQ ID NO: 30 or SEQ ID NO: 60, wherein the C-terminus may be amidated. In some embodiments, the N-terminus of the OATP-binding peptide may be modified with a hydrophobic group, and / or the C-terminus may be modified with other modifications. In some embodiments, the N-terminus of the OATP-binding peptide may be modified with a hydrophobic group, and / or the C-terminus may be amidated. In certain embodiments, the OATP-binding peptide may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, wherein the N-terminus may be modified with a hydrophobic group, and / or the C-terminus may be amidated. For example, the OATP-binding peptide may comprise the amino acid sequence set forth in SEQ ID NO: 30 or SEQ ID NO: 60, with its N-terminus modified with a hydrophobic group selected from the group consisting of myristic acid (myr), palmitic acid (plam), stearic acid (stearoyl), or cholesterol (chol), and its C-terminus modified with amidation, such as the polypeptides set forth in SEQ ID NOs: 67-71. In some embodiments, the OATP-binding peptide may be modified with a hydrophobic group selected from the group consisting of myristic acid (plam), and / or amidation at the C-terminus. In certain embodiments, the polypeptide may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, with its N-terminus modified with myristoylation and / or amidation at the C-terminus.For example, the OATP-binding peptide may comprise a polypeptide as shown in SEQ ID NO: 30, 36-42 or 60, wherein the N-terminus thereof is modified with a myristic acid (myr) hydrophobic group and the C-terminus thereof is amidated, as shown in any one of SEQ ID NO: 67, 68 and 72-78.

[0141] The OATP-binding peptides and variants thereof derived from hepadnaviruses provided herein retain at least the biological activity of binding to OATP and can be used to prevent and / or treat hepadnavirus and / or HDV infection and related diseases. In some embodiments, the OATP-binding peptides are derived from, but are not limited to, orthohepadnavirus, avihepadnavirus, metahepadnavirus, herpetohepadnavirus, and other hepadnaviruses. In some embodiments, the OATP-binding peptide is derived from, but is not limited to, human (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkey, woodchuck hepatitis virus (WHV), arctic shrew, ground shrew (GSHV), tree shrew, chimpanzee, bat (BHV) and other animal hepadnaviruses, and Avihepadnavirus that infects duck (DHBV), heron (HHBV) and other birds. In some embodiments, the OATP-binding peptide is derived from, but is not limited to, the Pre-S region of the HBV surface antigen of genotypes A, B, C, D, E, F, G, H and I. In some embodiments, the OATP-binding peptide is derived from, but is not limited to, amino acids 13-59 of genotype C HBV Pre-S. The N-terminus of the OATP-binding peptide is modified with a hydrophobic group, and the C-terminus may be modified with a stabilizing group or deleted. In some embodiments, the N-terminus of the OATP-binding peptide is modified with a hydrophobic group such as myristic acid (myr), palmitic acid (plam), stearic acid (stearoyl), or cholesterol (chol), and the C-terminus may be amidated. In some embodiments, the N-terminus of the OATP-binding peptide is modified with a hydrophobic group such as myristic acid (myr), and the C-terminus may be amidated. The OATP-binding peptide retains at least the biological activity of binding to OATP, particularly retaining the biological activity of binding to OATP1B1, and / or OATP1B2, and / or OATP1B3, and particularly retaining the biological activity of binding to human OATP1B1 and / or human OATP1B3. In some embodiments, the OATP-binding peptide structure derived from the hepadnavirus is a polypeptide and variants thereof as shown in any of the exemplary structures of SEQ ID NOs: 67-78, particularly including but not limited to polypeptides and variants thereof having the exemplary structures of SEQ ID NOs: 67 and 68.

[0142] In some embodiments, the OATP-binding peptides can be used to prevent and / or treat hepadnavirus and / or HDV infection and related diseases, particularly for the prevention and treatment of HBV and / or HDV infection and related diseases in humans. The diseases include, but are not limited to, acute or chronic HBV and / or HDV infection, acute viral hepatitis B and / or D, chronic viral hepatitis B and / or D, fulminant viral hepatitis B and / or D, subsevere viral hepatitis B and / or D, severe viral hepatitis B and / or D, HBV and / or HDV carrier infection, HBV and / or HDV infection-related liver disease, HBV and / or HDV infection-related liver fibrosis, HBV and / or HDV infection-related liver cirrhosis, HBV and / or HDV infection-related liver cancer, HBV and / or HDV infection-related liver function abnormalities, HBV and / or HDV infection-related liver function decompensation, HBV and / or HDV infection-related liver failure, and other related diseases. The methods for treating or preventing liver disease described herein are methods for treating or preventing HBV and / or HDV infection and related diseases, including but not limited to administering a therapeutically effective or prophylactically effective amount of a hepadnavirus-derived peptide to a subject in need thereof, thereby preventing or reducing the interaction between the hepadnavirus and / or HDV and OATP, thereby blocking or inhibiting infection by the hepadnavirus and / or HDV. For example, but not limited to, in the event of accidental exposure to HBV and / or HDV, the OATP-binding peptide described herein can be administered to block viral entry into liver cells through binding of the OATP-binding peptide to the OATP, thereby blocking viral infection and achieving disease prevention. In some embodiments, provided herein are methods for blocking HBV and / or HDV infection of liver tissue or liver cells and / or blocking HBV and / or HDV spread between liver cells, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition of an OATP-binding peptide derived from HBVPre-S1. In some embodiments, the disease is a liver disease associated with HBV and / or HDV infection. By using the various OATP-binding peptides derived from HBV Pre-S1 described herein to bind to OATP, the virus can be blocked from entering cells via OATP, thereby preventing or reducing viral infection of liver cells and blocking the onset and / or progression of the disease. In some embodiments, this document also provides the use of OATP-binding peptides in the preparation of medicaments for treating or preventing liver diseases, particularly liver diseases associated with hepatitis B virus or hepatitis D virus infection, as well as the use of OATP-binding peptides in the preparation of medicaments for blocking HBV and / or HDV infection of liver tissue or liver cells and / or blocking the spread of the viruses between liver cells.In some embodiments, the OATP-binding peptides described herein can be used in combination with HBV and / or HDV therapeutic or preventive drugs, including but not limited to nucleoside (acid) analogs (including but not limited to lamivudine, telbivudine, entecavir, tenofovir disoproxil, adefovir dipivoxil, clavudine, tenofovir disoproxil and other nucleoside (acid) analogs), interferons (including but not limited to conventional interferons (including but not limited to interferon α-2a, α-2b and other types of interferons), long-acting interferons, PEG interferons (including but not limited to PEG interferon α-2a, α-2b and other types of PEG interferons) and other interferons), nucleocapsid inhibitors, RNA interference, immunomodulators and other drugs for preventing or treating HBV or HDV infection, as well as combined use regimens of these drugs.

[0143] HBV and / or HDV infection model and its establishment method

[0144] Also provided herein is a cell or animal model susceptible to HBV and / or HDV infection and a method for establishing the same. The method comprises transfecting a cell or animal with an exogenous SLCO gene and / or expressing an exogenous OATP protein in the cell or animal, thereby rendering the cell or animal, which was previously non-susceptible or not susceptible to infection with HBV and / or HDV, susceptible to HBV and / or HDV after transfection with the SLCO gene or expression of the exogenous OATP protein. Also provided herein are drugs and lead compounds screened using the aforementioned HBV and / or HDV-infected cell and / or animal models, wherein the drugs are used to treat and / or prevent HBV and / or HDV infection. The method for establishing a cell or animal model susceptible to HBV and / or HDV infection comprises constructing a cell or animal model susceptible to HBV and / or HDV infection by transfecting the cell or animal with an exogenous SLCO gene and / or expressing an exogenous OATP protein. The cells or animals are not susceptible or not easily infected by HBV and / or HDV before being transfected with the exogenous SLCO gene and / or expressing the exogenous OATP protein.

[0145] The exogenous SLCO gene and exogenous OATP protein are derived from, but are not limited to, mammals, birds, fish, amphibians, and other animals. The mammals include, but are not limited to, primates, tree shrews (tupaia), rodents, bats, even-toed ungulates (Artiodactyla), Carnivora, and other mammals. The birds include, but are not limited to, ducks, herons, and other avian animals. The primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. The hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora. In some embodiments herein, the exogenous SLCO gene and OATP protein are derived from mammals or birds.In some preferred embodiments herein, the exogenous SLCO gene and OATP protein are derived from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic hamsters, ground shrews, bats, woolly monkeys, capuchin monkeys, and squirrel monkeys. In some preferred embodiments herein, the exogenous SLCO gene and OATP protein are derived from humans, chimpanzees, and tree shrews. In some further preferred embodiments herein, the transfected exogenous SLCO gene and OATP protein are derived from humans.

[0146] The exogenous SLCO genes include, but are not limited to, SLCO1, SLCO2, SLCO3, SLCO4, SLCO5, SLCO6, SLCO7, SLCO8, SLCO9, SLCO10, SLCO11, and other SLCO gene families. In some embodiments, the SLCO genes include, but are not limited to, the SLCO gene family expressed in hepatocytes. In some embodiments, the SLCO genes include, but are not limited to, the SLCO1B subfamily. In some preferred embodiments, the SLCO genes include, but are not limited to, the SLCO1B subfamily expressed in hepatocytes. In some preferred embodiments, the SLCO genes include, but are not limited to, SLCO1B3, SLCO1B1, SLCO1B2, and other members of the SLCO1B gene subfamily. In some embodiments herein, the transfected exogenous SLCO genes are SLCO1B3 and / or SLCO1B1 and / or SLCO1B2, or a combination of any two thereof, or SLCO1B3, SLCO1B1, and SLCO1B2. In some embodiments herein, the transfected exogenous SLCO1B3 gene is derived from, but is not limited to, humans (see SEQ ID NO: 3 for an exemplary sequence, NCBI Reference Sequence: NM_019844), chimpanzees (see genebank Gene ID: 749254 for an exemplary sequence), tree shrews (see genebank Gene ID: 102476084 for an exemplary sequence), cynomolgus monkeys (see genebank Gene ID: 102136593 for an exemplary sequence), Arctic ground hamsters (genebank Gene ID: 113185010), pigs (see genebank Gene ID: 100620829 for an exemplary sequence), dogs (see genebank Gene ID: 403519 for an exemplary sequence), and variants thereof.and SLCO1B3 genes and variants thereof from other animals mentioned above, including but not limited to gene banks NC_050564, NC_013676, NW_017869963, NW_006501111, NW_022196903, NW_003613944, NW_004801640, NW_004949097, NW_015351262, NW_024404945, NW_020539983, NW_023144717, NW_006711288, NW_019154091, NW_004569192, NC _044309, NW_020339853, NC_018729, NW_017619867, NW_023365217, NC_051831, NW_020656135, NW_022631003, NC _045619, NW_004451484, NC_043709, NW_020356453, NC_048233, NW_020312893, NW_020998839, NW_006383153, NW_ 023270504, NC_032654, NC_010447, NW_006781351, NW_023601653, NW_011889225, NC_009149, NW_007675154, NW_ 023458025, NW_006159939, NW_023666069, NW_012009027, NC_022282, NC_041764, NW_012010756, NC_044984, NC_0 37679, NW_012109719, NW_022681456, NW_016809129, NC_044558, NC_045443, NC_044614, NC_048251, NC_036891, NC_000012, NC_036915, NC_044403, NW_022611650, NW_024100944, NC_033666, NW_006408616 and other SLCO1B3 gene sequences and variants thereof. In some embodiments herein, the transfected exogenous SLCO1B1 gene originates from, but is not limited to, humans (see SEQ ID NO: 6, Gene ID: 10599 for an exemplary sequence), bonobos (see Gene ID: 100976318 for an exemplary sequence), cynomolgus monkeys (see Gene ID: 102137347 for an exemplary sequence), and variants thereof.and SLCO1B1 gene sequences and variants thereof from other animals mentioned above, including but not limited to gene banks NC_000012, NC_041764, NC_036891, NC_022282, NC_048391, NW_012009027, NW_012014011, NC_037679, NW_012109719, NC_044614, NC_048251, NC_036915, NW_006399825, NW_023666069, NW_016809129, NC_044558, NC_044403, NW_022611650, NW_012112030, NC_045443, NW_006399825 and other SLCO1B1 gene sequences and variants thereof. In some preferred embodiments, the transfected exogenous SLCO gene is derived from human SLCO1B3 (see SEQ ID NO: 3, NCBI Reference Sequence: NM_019844 for an exemplary sequence), chimpanzee SLCO1B3 (see genebank Gene ID: 749254 for an exemplary sequence), tree shrew SLCO1B3 (see genebank Gene ID: 102476084 for an exemplary sequence), and / or human SLCO1B1 (see SEQ ID NO: 6, genebank Gene ID: 10599 for an exemplary sequence), chimpanzee, or tree shrew. In some further preferred embodiments, the transfected exogenous SLCO gene is derived from human SLCO1B3 (see SEQ ID NO: 3, NCBI Reference Sequence: NM_019844) and / or human SLCO1B1 (see SEQ ID NO: 6, genebank Gene ID: 10599 for an exemplary sequence). The SLCO gene includes, but is not limited to, the complete genome sequence of SLCO and its variants, coding sequence (CDS) and its variants, open reading frame sequence (ORF) and its variants, and other forms of gene coding sequence. In some embodiments herein, the transfected exogenous SLCO gene also includes variants, fragments, and homologous sequences of the above-mentioned exogenous SLCO gene sequence. Also included herein are vectors carrying the above-mentioned SLCO gene and its variants, fragments, and homologous sequences, including but not limited to DNA, RNA, plasmids, viruses, and other vectors.The methods for transfecting the exogenous SLCO gene include but are not limited to liposomes, cationic peptides, electrofection, viral vectors, transgenics, gene insertion, gene editing and other methods that can transfect the exogenous SLCO gene into cells or animals. The technical feature is that DNA or RNA carrying the SLCO gene coding is transferred into cells or animals.

[0147] The exogenous OATP proteins include but are not limited to OATP1, OATP2, OATP3, OATP4, OATP5, OATP6, OATP7, OATP8, OATP9, OATP10, OATP11 and other OATP families. In some embodiments, the OATP family includes but is not limited to the OATP family expressed in hepatocytes. In some embodiments, the OATP includes but is not limited to the OATP 1B subfamily. In some preferred embodiments, the OATP includes but is not limited to the OATP 1B subfamily expressed in hepatocytes. In some preferred embodiments, the OATP includes but is not limited to OATP1B3, OATP1B1, OATP1B2 and other OATP1B subfamily members. In some embodiments herein, the transfected exogenous OATP is OATP1B3 and / or OATP1B1 and / or OATP1B2, or a combination of two thereof, or OATP1B3, OATP1B1 and OATP1B2. In some embodiments herein, the exogenous OATP1B3 protein expressed is derived from, but is not limited to, humans (an exemplary amino acid sequence can be found in SEQ ID NO: 1, an exemplary coding sequence can be found in SEQ ID NO: 2, NCBI Reference Sequence: NM_019844), chimpanzee (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_016923821), tree shrew (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006155085), cynomolgus monkey (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001283191), Arctic ground squirrel (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_026391932), pig (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001315607), dog (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001166047), and OATP1B3 proteins and variants thereof derived from other animals mentioned above.In some embodiments herein, the expressed exogenous OATP1B1 protein is derived from, including but not limited to, humans (an exemplary amino acid sequence can be found in SEQ ID NO: 4, an exemplary coding sequence can be found in SEQ ID NO: 5, NCBI Reference Sequence: NM_006446), bonobos (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003828901), cynomolgus monkeys (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001284540), and OATP1B1 proteins and variants thereof derived from the above-mentioned other animals. In some preferred embodiments, the exogenous OATP protein expressed is derived from human OATP1B3 (an exemplary amino acid sequence can be found in SEQ ID NO: 1, an exemplary nucleic acid coding sequence can be found in SEQ ID NO: 2, NCBI Reference Sequence: NM_019844), chimpanzee OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_016923821), tree shrew OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006155085), and / or human OATP1B1 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_006446), chimpanzee OATP1B1 or tree shrew OATP1B1. In some further preferred embodiments, the exogenous OATP protein expressed is derived from human OATP1B3 (an exemplary amino acid sequence can be found in SEQ ID NO: 1, an exemplary nucleic acid coding sequence can be found in SEQ ID NO: 2, NCBI Reference Sequence: NM_019844) and / or human OATP1B1 (an exemplary amino acid sequence can be found in SEQ ID NO: 4, an exemplary nucleic acid coding sequence can be found in SEQ ID NO: 5, NCBI Reference Sequence: NM_006446). In some embodiments herein, the exogenous OATP protein expressed also includes variants, fragments, and homologous sequences of the amino acid sequences, and variants, fragments, and homologous sequences of the nucleic acid coding sequences of the above-mentioned exogenous OATP proteins. The methods for expressing the exogenous OATP protein include, but are not limited to, incubating the OATP protein or recombinant protein with cells, or performing cell fusion with cells expressing the OATP protein, or coupling the OATP protein to the cell surface, or introducing a DNA or RNA fragment or vector containing the nucleic acid encoding the OATP protein into cells or animals, as well as other methods for causing cells or animals to express the exogenous OATP protein.

[0148] The cell or animal model susceptible to HBV and / or HDV and its establishment method can be transfected with an exogenous SLCO gene alone or simultaneously with an exogenous SLC10A1 gene; or can express an exogenous OATP protein alone or simultaneously with an exogenous NTCP protein. The exogenous SLC10A1 gene and exogenous NTCP protein are derived from, but are not limited to, mammals, birds, fish, amphibians, and other animals. The mammals include, but are not limited to, primates, tree shrews (tupaia), rodents, bats, even-toed ungulates (Artiodactyla), Carnivora, and other mammals. The birds include, but are not limited to, ducks, herons, and other avian animals. The primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. The hominoid species include but are not limited to humans, chimpanzees, gorillas, gibbons, orangutans and other hominoid species. The New World monkeys include but are not limited to woolly monkeys, capuchin monkeys, squirrel monkeys and other New World monkeys. The Old World monkeys include but are not limited to crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include but are not limited to Sciuridae animals, myomorpha and other rodents. The Sciuridae animals include but are not limited to arctic ground squirrels, ground squirrels, woodchucks and other Sciuridae animals. The myomorpha animals include but are not limited to rats (Rattus rattus),Mice (Mus musculus) and other rodents (myomorpha). The even-toed ungulate (Artiodactyla) animals include but are not limited to pigs and other even-toed ungulates. The Carnivora animals include but are not limited to dogs and other Carnivora animals. In some embodiments herein, the exogenous SLC10A1 gene and NTCP protein are derived from mammals or birds. In some preferred embodiments herein, the exogenous SLC10A1 gene and NTCP protein are derived from humans, chimpanzees, bonobos, tree shrews, marmots, Arctic shrews, ground shrews, bats, woolly monkeys, capuchins, squirrel monkeys, rats, and mice. In some preferred embodiments herein, the exogenous SLC10A1 gene and NTCP protein are derived from humans, chimpanzees, and tree shrews. In some further preferred embodiments herein, the transfected exogenous SLC10A1 gene and NTCP protein are derived from humans. In some embodiments herein, the exogenous SLC10A1 gene transfected simultaneously with the exogenous SLCO gene is derived from, but is not limited to, humans (see SEQ ID NO: 79, GeneID: 6554 for an exemplary sequence), chimpanzees (see GeneID: 453001 for an exemplary sequence), tree shrews (see GeneID: 102494 for an exemplary sequence), bonobos (see GeneID: 100985481 for an exemplary sequence), capuchins (see GeneID: 116528519 for an exemplary sequence), squirrel monkeys (see GeneID: 101045215 for an exemplary sequence), cynomolgus monkeys (see GeneID: 102138916 for an exemplary sequence), arctic ground hamsters (see GeneID: 113180315 for an exemplary sequence), ground hamsters (see GeneID: 113180315 for an exemplary sequence), rats (see GeneID: 116905231 for an exemplary sequence), and mice (see GeneID: 116905231 for an exemplary sequence). GeneID: 20493), pig (example sequence see genebank GeneID: 100153302), dog (example sequence see genebank GeneID: 480372), and SLC10A1 genes and variants thereof from the above-mentioned other animals, as well as SLC10A1 genes and variants thereof from the above-mentioned other animals, including but not limited to the following example sequences see: gene_bank_NC_000014.9, NC_051341.1, NC_000078.7, NC_041760.1, NC_037337.1, NC_007131.7, NC_030684.2, NC_036893.1,NC_051812.1, NC_048392.1, NC_008801.1, NW_004569144.1, NC_044068.1, NW_024429186.1, NC_048231.1, NC_010449.5, NW_005395103.1, NW_017364105.1, NC_009167.3, NW_019174280.1, NC_050533.1, NC_035899.1, NC_019880.2, NW_006890081.1, NW_005819088.1, NC_023185.1, NC_045166. 1、NC_030440.1、NC_049537.1、NC_047591.1、NC_036849.1、NC_046375.1、NC_024351.1、NC_036457.1、NC_045686.1、NW_005179644.1、NC_039316.1、NC_041348.1、NC_044208.1、NW_017859650.1、NC_051245.1、NW_007281450.1、NW_017713697.1、NW_017728947.1、NC_048605.1、NW_020451983.1、 NC_051776.1, NW_023336592.1, NC_048897.1, NW_009952265.1, NW_004973262.1, NC_034414.1, NC_052536.1, NW_022044854.1, NC_031774.1, NW_014650483.1, NW_009152437.1, NW_008795283.1, NW_008824949.1, NC_041728.1, NC_045427.1, NC_050580.1, NW_004491405.1, NW_006804717.1, N W_022103915.1, NW_006438074.1, NW_023416306.1, NW_023537909.1, NW_020093309.1, NW_022436999.1, NW_012007369.1, NC_022278.1, NW_012015021.1, NC_044982.1, NC_037675.1, NW_012106931.1, NC_044616.1, NC_048253.1, NC_036917.1, NW_020356506.1, NC_018728.3, NW_017619866.1,NW_020339215.1、NC_045600.1、NW_004450335.1、NW_023399451.1、NW_006383727.1、NW_023270825.1、NW_022589710.1、NW_022134863.1、NW_004438415.1、NC_047035.1、NW_ 022098036.1, NC_041224.1, NC_045786.1, NW_003573429.1, NW_014638379.1, NW_007676261.1, NW_011515408.1, NC_044516.1, NW_018335685.1, NC_032659.1, NC_030817.1, NC_040258.1, NW_023435897.1, NC_050555.1, NW_003159375.1, NW_015351211.1, NW_020539924.1, NW_012267218.1, NW_003613686.1, NC_051075.1, NW_018657848.1, NC_034 581.1、NC_034596.1、NC_046160.1、NT_176399.1、NW_004524583.1、NW_004624734.1、NW_005857041.1、NW_023425344.1、NW_024100921.1、NW_003338865.1、NW_006399787.1、 NW_015150892.1、NW_024425008.1、NW_007370659.1、NW_018734275.1、NC_044498.1、NW_009900513.1、NW_020455250.1、NC_040085.1、NW_021964165.1、NC_033662.1、NW_003852510.1、NC_042286.1、NC_048301.1、NC_040242.1、NW_020834 750.1, NW_004955466.1, NW_020312827.1, NW_020998607.1, NW_022196912.1, NC_045618.1, NC_051448.1, NC_043708.1, NW_018503592.1, NW_018344057.1, NW_005842135.1, NC_050456.1, NW_021703778.1, NC_045764.1, NW_004545910.1,NC_051214.1, NW_024408640.1, NW_011494811.1, NW_018127943.1, NW_005081537.1, NC_052956.1, NW_006799940.1, NW_015112671.1, NW_015095634.1, NW_005054301.1, NW_009646445.1, NW_023601104.1, NW_023416313.1, NW_004504313.1, NW_017870438.1, NW_010972853.1, NW_022681468. 1、NC_045750.1、NW_009632203.1、NW_022665528.1、NC_041378.1、NW_018114435.1、NW_008609214.1、NW_005871133.1、NW_023425404.1、NC_046289.1、NC_045004.1、NC_021677.1、NW_023666053.1、NW_023044993.1、NC_044308.2、NW_016820119.1、NC_044553.1、NC_044381.1、NC_046632.1、NC_ 041312.1, NW_006712282.1, NW_015505194.1, NW_022631221.1, NC_022009.1, NC_046312.1, NW_019029691.1, NW_019372207.1, NW_022611658.1, NC_050482.1, NW_008239553.1, NC_044575.1, NC_037555.1, NC_040903.2, NW_020837954.1, NC_046226.1, NW_023144640.1, NW_010582752.1, NW_0 17729025.1, NW_010537981.1, NW_010771612.1, NW_018150791.1, NW_022641422.1, NW_017805708.1, NW_005369967.1, NC_041820.1, NW_020656171.1, NW_006781042.1, NW_010035470.1, NW_009002855.1, NW_011888796.1, NW_023455365.1, NW_004567114.1, NW_007577740.1, NW_015164181.1,NW_019776417.1, NC_040901.1, NC_042532.1, NW_010423023.1, NW_009928727.1, NW_005087567.1, NC_043759.1, NW_006408597.1 , NW_009874831.1, NW_009258894.1, NW_023528387.1, NW_020799101.1, NW_015090839.1, NW_014006077.1, NC_046656.1, NC_04104 3.1. 1. NW_018030485.1, NC_047090.1, NC_050191.1, NW_023365123.1, NW_015505036.1, NC_042571.1, NC_041720.1, NW_020448008.1, N W_006727797.1, NW_012116265.1, NC_043979.1, NW_012150840.1, NW_013185668.1, NC_045964.1, NC_045701.1, NC_041066.1, NW_ 009270392.1, NW_007729665.1, NW_021604585.1, NC_044003.1, NC_045439.1, NC_046335.1, NW_008348666.1, NC_045481.1, NW_006 921675.1, NW_020848012.1, NC_044039.1, NW_015553274.1, NW_019715876.1, NW_020172672.1, NC_044325.1, NW_019525097.1, NW_007014397.1, NC_046358.1, NW_020664447.1, NW_016107905.1, NC_047530.1, NW_004454173.1, NW_004443946.1, NW_019154104.1 and other SLC10A1 gene sequences and variants thereof. In some preferred embodiments, the transfected exogenous SLC10A1 gene is derived from human (see SEQ ID NO: 79, genebank GeneID: 6554 for an exemplary sequence),Chimpanzee (example sequence see genebank GeneID: 453001), tree shrew (example sequence see genebank GeneID: 102494). In some more preferred embodiments, the transfected exogenous SLC10A1 gene is derived from human (example sequence see SEQ ID NO: 79, genebank GeneID: 6554). The SLC10A1 gene includes but is not limited to the complete genome sequence of SLC10A1 and its variants, coding sequence (CDS) and its variants, open reading frame sequence (ORF) and its variants, and other forms of gene coding sequence. In some embodiments herein, the transfected exogenous SLC10A1 gene also includes variants, fragments, and homologous sequences of the above-mentioned exogenous SLC10A1 gene sequence. The present invention also includes vectors carrying the above-mentioned SLC10A1 gene and its variants, fragments, and homologous sequences, including but not limited to DNA, RNA, plasmids, viruses, and other vectors. The method of transfecting the exogenous SLC10A1 gene includes but is not limited to the use of liposomes, cationic peptides, electrofection, viral vectors, transgenics, gene insertion, gene editing and other methods that can transfect the exogenous SLC10A1 gene into cells or animals. The technical feature is that DNA or RNA encoding the SLC10A1 gene is transferred into cells or animals.

[0149] In some embodiments herein, the exogenous NTCP protein expressed simultaneously with the exogenous OATP protein is derived from, but is not limited to, humans (an exemplary amino acid sequence can be found in SEQ ID NO: 7, an exemplary nucleic acid coding sequence can be found in SEQ ID NO: 80, NCBI Reference Sequence: NM_003049), chimpanzee (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_510035), tree shrew (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006171503), bonobo (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003824101), capuchin monkey (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_032245781), squirrel monkey (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003924480), cynomolgus monkey (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003924481), and cynomolgus monkey (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003924482). Sequence: NM_001283323), Arctic hamster (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_026385176), hamster (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_005322843), rat (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_032908052), mouse (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001177561), pig (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_001927695), dog (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_537494), as well as NTCP and variants thereof derived from the above-mentioned other animals.

[0150] In some preferred embodiments, the exogenous NTCP protein expressed is derived from humans (an exemplary amino acid sequence can be found in SEQ ID NO: 7, an exemplary nucleic acid coding sequence can be found in SEQ ID NO: 80, NCBI Reference Sequence: NM_003049), chimpanzee (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_510035), or tree shrew (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006171503). In some further preferred embodiments, the exogenous NTCP protein expressed is derived from humans (an exemplary amino acid sequence can be found in SEQ ID NO: 7, an exemplary nucleic acid coding sequence can be found in SEQ ID NO: 80, NCBI Reference Sequence: NM_003049). In some embodiments herein, the expressed exogenous NTCP protein also includes variants, fragments, and homologous sequences of the amino acid sequences of the exogenous NTCP proteins described above. The method for expressing the exogenous NTCP protein includes but is not limited to incubating the NTCP protein with cells, or coupling the NTCP protein to the cell surface, or introducing a DNA or RNA fragment or vector containing a coding sequence encoding the NTCP protein into cells or animals, as well as other methods that can cause cells or animals to express the exogenous NTCP protein.

[0151] In some embodiments herein, cells transfected with an exogenous SLCO gene and / or expressing an exogenous OATP protein include, but are not limited to, human hepatocyte cell lines, mouse hepatocyte cell lines, rat hepatocyte cell lines, and other animal-derived primary hepatocytes or hepatocyte cell lines. Human hepatocyte cell lines include, but are not limited to, HepG2, Huh7, Hep RG, SMMC-7721, Bel-7402, MHCC97, Hep3B, PLC / PRF / 5, HepG2.2.15, Hep AD38, HL-7702, QSG-7701, L-02, HL-7702BaPT, HCC-DJ711, THLE-3, HC-04, and other human liver-derived cell lines. Mouse hepatocyte lines include, but are not limited to, GHA1, AML12, BNL CL.2, NCTC 1469, NCTC 1469, FL83B, H2.35, Hepa1-6, Hep56.1D, and other mouse hepatocytes. Rat hepatocyte lines include, but are not limited to, IAR20, H-4-II-E, H4-II-E-C3, BRL, BRL 3A, TC5123, and other rat hepatocytes. Woodchuck hepatocyte lines include, but are not limited to, WCH-17 and other woodchuck hepatocytes. Tree shrew hepatocyte lines include, but are not limited to, ITH48, ITH6.1, and other tree shrew hepatocytes. Rabbit hepatocyte lines include, but are not limited to, RNH / HL-034 and other rabbit hepatocytes. Porcine hepatocyte lines include, but are not limited to, PNH / HL-059 and other porcine hepatocytes. Hepatocytes from other animal sources are also encompassed herein. In some preferred embodiments, the cells are selected from, but are not limited to, human hepatocyte cell lines HepG2, Huh7, Hep RG, mouse hepatocyte cell lines Hepa1-6, Hep56.1D, or rat hepatocyte cell line TC5123. In some further preferred embodiments, cells transfected with an exogenous SLCO gene and / or expressing an exogenous OATP protein include, but are not limited to, human hepatocyte cell lines HepG2 or Huh7. In some embodiments herein, animals transfected with an exogenous SLCO gene and / or expressing an OATP protein include, but are not limited to, mammals, birds, and other animals. Mammals include, but are not limited to, mice, rats, dogs, monkeys, tree shrews, pigs, rabbits, marmots, tree shrews, and other mammals. Birds include, but are not limited to, ducks, herons, and other birds. Other animals include bats and other animals.Mice include, but are not limited to, C57BL (including, but not limited to, C57BL / 6, C57BL / 10, and other strains), BALB / c (including, but not limited to, BALB / cd, BALB / cJ, BALB / cAnN, and other strains), C3H (including, but not limited to, C3H / Bi, C3H / He, C3H / HeJ, C3H / St, C3HeB / FeJ, C3H / DiSn, C3H / Sf, C3H / HeI, and other strains), DBA / 2 (including, but not limited to, DBA / 2, DBA / 2J, and other strains), CBA (including, but not limited to, CBA / Br, CBA / Ca, CBA / J, CBA / st, CBA / H, and other strains), A (including, but not limited to, A / He, A / J, A / WySN, and other strains), AKR (including, but not limited to, AKR / A, AKR / J, AKR / N, and other strains), Kunming mice, golden hamsters, NIH albino mice, ICR mice, Webster mice, LACA mice, and other mice. Rats include, but are not limited to, Wistar, SD, ACI, F344, LEW, LOU / CN, LOU / MN, SHR, WKY, GH, Long-Evans, Brown Norway, and other rats. Dogs include, but are not limited to, Beagle, Chinese domestic dog, and other dogs. Monkeys include, but are not limited to, cynomolgus macaques, rhesus macaques, red-faced macaques, bear macaques, and other monkeys. Rabbits include, but are not limited to, Chinese rabbits, Chinchilla rabbits, Big-eared White rabbits, New Zealan white rabbits, Silver Fox rabbits, Himalayan albio rabbits, Angora rabbits, Belgian Hare rabbits, Lop rabbits, California rabbits, Chekered Giant rabbits, Danish White rabbits, West German Angora rabbits, and other rabbits. In some preferred embodiments herein, animals transfected with exogenous SLCO genes and / or expressing exogenous OATP proteins include, but are not limited to, mice and rats. In a further preferred embodiment, animals transfected with exogenous SLCO gene and / or expressing exogenous OATP protein include but are not limited to C57BL mice, BALB / c mice, SD rats, and Wistar rats.

[0152] This article also includes lead compounds or drugs for treating and / or preventing HBV and / or HDV infection and related diseases obtained by screening using the above-mentioned HBV and / or HDV susceptible cells and / or animal models, including but not limited to small molecule drugs, proteins, peptides, antibodies, poly RNA, poly DNA and other drugs. The lead compounds or drugs can be used to include but are not limited to blocking or inhibiting HBV and / or HDV infection, blocking or inhibiting HBV and / or HDV replication, clearing HBV and / or HDV infection, blocking or inhibiting HBV and / or HDV release, activating and / or regulating HBV and / or HDV specific immune responses, and other methods for treating and / or preventing HBV and / or HDV infection and related diseases. In some embodiments herein, the HBV and / or HDV susceptible cells and / or animal models described herein are used to screen a drug library to obtain lead compounds or drugs that can be used to treat and / or prevent HBV and / or HDV infection and related diseases. The drug library includes but is not limited to a small molecule compound library, a protein library, an antibody library, a peptide library, a natural product library, and other drug or lead structure libraries. In some embodiments herein, the HBV and / or HDV susceptible cells and / or animal models described herein are used to obtain drug targets for the life cycle of HBV and / or HDV, including infection, replication, and progeny virus release, and small molecule drugs, proteins, peptides, antibodies, polyRNAs, polyDNAs, and other drugs are designed for these targets, or small molecule compound libraries, protein libraries, antibody libraries, peptide libraries, natural product libraries, and other agents or lead structure libraries are screened using these targets to obtain lead compounds or drugs that can be used to treat and / or prevent HBV and / or HDV infection and related diseases.

[0153] A drug delivery method and carrier targeting OATP

[0154] Provided herein is a method and carrier for delivering a drug or group, wherein the method delivers the drug or group by targeting or binding to OATP, and the carrier at least retains the biological activity of targeting or binding to OATP and delivering the carried drug or group into cells or organs.

[0155] The OATPs targeted or bound by the delivery methods and vectors include, but are not limited to, OATPs from mammals, birds, fish, amphibians, and other animals. The mammals include, but are not limited to, primates, tree shrews, rodents, bats, artiodactyla, carnivora, and other mammals. The birds include, but are not limited to, ducks, herons, and other avian animals. The primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. The hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora. In some embodiments herein, the OATP targeted by the vector is a mammalian or avian OATP.In some preferred embodiments herein, the OATP targeted by the vector is human, chimpanzee, bonobo, tree shrew, marmot, Arctic shrew, ground shrew, bat, woolly monkey, capuchin monkey and squirrel monkey OATP. In some preferred embodiments herein, the OATP targeted by the vector is human, chimpanzee, tree shrew OATP. In some further preferred embodiments herein, the OATP targeted by the vector is human OATP.

[0156] The OATPs targeted or bound by the delivery methods and vectors include, but are not limited to, OATP 1, OATP2, OATP3, OATP4, OATP5, OATP6, OATP7, OATP8, OATP9, OATP10, OATP11, and other OATP families. In some embodiments, the OATPs targeted by the delivery methods and vectors include, but are not limited to, the OATP family expressed in hepatocytes. In some embodiments, the OATPs targeted by the delivery methods and vectors include, but are not limited to, the OATP1B subfamily. In some preferred embodiments, the OATPs targeted by the delivery methods and vectors include, but are not limited to, the OATP1B subfamily expressed in hepatocytes. In some preferred embodiments, the OATPs targeted by the delivery methods and vectors include, but are not limited to, OATP1B3, OATP1B1, OATP1B2, and other OATP1B subfamily members. In some embodiments herein, the OATPs targeted by the delivery methods and vectors are OATP1B3 and / or OATP1B1 and / or OATP1B2, or a combination of any two thereof, or OATP1B3, OATP1B1, and OATP1B2. In some embodiments herein, the OATP targeted by the delivery methods and vectors includes, but is not limited to, human OATP1B3 (an exemplary amino acid sequence can be found in SEQ ID NO: 1, NCBI Reference Sequence: NM_019844), chimpanzee OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_016923821), tree shrew OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006155085), cynomolgus monkey OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001283191), Arctic hamster OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_026391932), porcine OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001315607), dog OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006155085), Sequence: NM_001166047), as well as OATP1B3 of the other animals mentioned above.In some embodiments herein, the OATP targeted by the delivery method and vector includes but is not limited to human OATP1B1 (an exemplary amino acid sequence can be found in SEQ ID NO: 5, NCBI Reference Sequence: NM_006446), bonobo OATP1B1 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003828901), cynomolgus monkey OATP1B1 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001284540), and OATP1B1 of the other animals mentioned above. In some preferred embodiments, the OATP targeted by the delivery method and vector is human OATP1B3 (an exemplary amino acid sequence can be found in SEQ ID NO: 1, NCBI Reference Sequence: NM_019844), chimpanzee OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_016923821), tree shrew OATP1B3 (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006155085), and / or human OATP1B1 (an exemplary amino acid sequence can be found in SEQ ID NO: 4, NCBI Reference Sequence: NM_006446), chimpanzee OATP1B1 or tree shrew OATP1B1. In some further preferred embodiments, the OATP targeted by the delivery methods and vectors is human OATP1B3 (an exemplary amino acid sequence can be found in SEQ ID NO: 1, NCBI Reference Sequence: NM_019844) and / or human OATP1B1 (an exemplary amino acid sequence can be found in SEQ ID NO: 4, NCBI Reference Sequence: NM_006446). In some embodiments herein, the OATP targeted by the delivery methods and vectors also includes variants, fragments, and homologous sequences of the amino acid sequences, and variants, fragments, and homologous sequences of the nucleic acid encoding sequences of the above-mentioned OATPs.

[0157] The delivery methods and vectors may target or bind to OATPs alone or simultaneously with OATPs and NTCPs. The delivery methods and vectors may simultaneously target or bind to NTCPs including, but not limited to, mammals, birds, fish, amphibians, and other animals. Mammals include, but are not limited to, primates, tree shrews (tupaia), rodents, bats, even-toed ungulates (Artiodactyla), Carnivora, and other mammalian NTCPs. Birds include, but are not limited to, ducks, herons, and other avian animals. Primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora.

[0158] In some embodiments herein, the NTCP to which the delivery method and vector simultaneously bind is mammalian or avian NTCP. In some preferred embodiments herein, the NTCP to which the delivery method and vector simultaneously bind is human, chimpanzee, bonobo, tree shrew, marmot, Arctic shrew, ground shrew, bat, woolly monkey, capuchin monkey, squirrel monkey, rat, and mouse NTCP. In some preferred embodiments herein, the NTCP to which the delivery method and vector simultaneously bind is human, chimpanzee, or tree shrew NTCP. In some further preferred embodiments herein, the NTCP to which the delivery method and vector simultaneously bind is human NTCP. In some embodiments herein, the OATP-targeted drug delivery method and OATP-targeted delivery vector, and the NTCP bound simultaneously include but are not limited to human NTCP (an exemplary amino acid sequence can be found in SEQ ID NO: 7, NCBI Reference Sequence: NM_003049), chimpanzee NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_510035), tree shrew NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006171503), bonobo NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003824101), capuchin NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_032245781), squirrel monkey NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003924480), cynomolgus monkey NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_003924481), and cynomolgus monkey NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006171503). Sequence: NM_001283323), Arctic hamster NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_026385176), hamster NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_005322843), rat NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_032908052), mouse NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_001177561), porcine NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_001927695),Canine NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_537494), as well as NTCP of other animals mentioned above and their variants, including but not limited to the following exemplary sequences: gene bank NC_000014.9, NC_051341.1, NC_000078.7, NC_041760.1, NC_037337.1, NC_007131.7, NC_030684.2, NC_036893.1, NC_051812.1, NC _048392.1, NC_008801.1, NW_004569144.1, NC_044068.1, NW_024429186.1, NC_048231.1, NC_010449.5, NW_005395103.1, NW_0173 64105.1, NC_009167.3, NW_019174280.1, NC_050533.1, NC_035899.1, NC_019880.2, NW_006890081.1, NW_005819088.1, NC_023185 .1, NC_045166.1, NC_030440.1, NC_049537.1, NC_047591.1, NC_036849.1, NC_046375.1, NC_024351.1, NC_036457.1, NC_045686.1, NW_005179644.1, NC_039316.1, NC_041348.1, NC_044208.1, NW_017859650.1, NC_051245.1, NW_007281450.1, NW_017713697.1, NW _017728947.1, NC_048605.1, NW_020451983.1, NC_051776.1, NW_023336592.1, NC_048897.1, NW_009952265.1, NW_004973262.1, N C_034414.1, NC_052536.1, NW_022044854.1, NC_031774.1, NW_014650483.1, NW_009152437.1, NW_008795283.1, NW_008824949.1, NC_041728.1, NC_045427.1, NC_050580.1, NW_004491405.1, NW_006804717.1, NW_022103915.1, NW_006438074.1, NW_023416306.1, NW_023537909.1、NW_020093309.1、NW_022436999.1、NW_012007369.1、NC_022278.1、NW_012015021.1、NC_044982.1、NC_037675.1、NW_012106931.1、NC_044616.1、NC_048253.1、NC_036917.1、NW_020356506.1、NC_018728.3、NW_017619866.1、NW_020339215.1、NC_045600.1、NW_004450335.1、NW_023399451.1、NW_ 006383727.1, NW_023270825.1, NW_022589710.1, NW_022134863.1, NW_004438415.1, NC_047035.1, NW_022098036.1, NC_041224.1, NC_045786.1, NW_003573429.1, NW_014638379.1, NW_007676261.1, NW_011515408.1, NC_044516.1, NW_018335685.1, NC_032659.1, NC_030817.1, NC_040258.1, NW_0 23435897.1, NC_050555.1, NW_003159375.1, NW_015351211.1, NW_020539924.1, NW_012267218.1, NW_003613686.1, NC_051075.1, NW_018657848.1, NC_034581.1, NC_034596.1, NC_046160.1, NT_176399.1, NW_004524583.1, NW_004624734.1, NW_005857041.1, NW_023425344.1, NW_024100921.1, NW _003338865.1、NW_006399787.1、NW_015150892.1、NW_024425008.1、NW_007370659.1、NW_018734275.1、NC_044498.1、NW_009900513.1、NW_020455250.1、NC_040085.1、NW_021964165.1、NC_033662.1、NW_003852510.1、NC_042286.1、NC_048301.1、NC_040242.1、NW_020834750.1、NW_004955466.1、NW_020312827.1, NW_020998607.1, NW_022196912.1, NC_045618.1, NC_051448.1, NC_043708.1, NW_018503592.1, NW_018344057.1, NW_005842135.1, NC_050456.1, NW_021703778.1, NC_045764.1, NW_004545910.1, NC_051214.1, NW_024408640.1, NW_011494811.1, NW_018127943.1, NW_00508153 7.1、NC_052956.1、NW_006799940.1、NW_015112671.1、NW_015095634.1、NW_005054301.1、NW_009646445.1、NW_023601104.1、NW_023416313.1、NW_004504313.1、NW_017870438.1、NW_010972853.1、NW_022681468.1、NC_045750.1、NW_009632203.1、NW_022665528.1、NC_041378.1、NW_018114435. 1、NW_008609214.1、NW_005871133.1、NW_023425404.1、NC_046289.1、NC_045004.1、NC_021677.1、NW_023666053.1、NW_023044993.1、NC_044308.2、NW_016820119.1、NC_044553.1、NC_044381.1、NC_046632.1、NC_041312.1、NW_006712282.1、NW_015505194.1、NW_022631221.1、NC_022009.1、NC_ 046312.1, NW_019029691.1, NW_019372207.1, NW_022611658.1, NC_050482.1, NW_008239553.1, NC_044575.1, NC_037555.1, NC_040903.2, NW_020837954.1, NC_046226.1, NW_023144640.1, NW_010582752.1, NW_017729025.1, NW_010537981.1, NW_010771612.1, NW_018150791.1, NW_022641422.1,NW_017805708.1、NW_005369967.1、NC_041820.1、NW_020656171.1、NW_006781042.1、NW_010035470.1、NW_009002855.1、NW_011888796.1、NW_023455365.1、NW_0 04567114.1、NW_007577740.1、NW_015164181.1、NW_019776417.1、NC_040901.1、NC_042532.1、NW_010423023.1、NW_009928727.1、NW_005087567.1、NC_043759.1、 NW_006408597.1, NW_009874831.1, NW_009258894.1, NW_023528387.1, NW_020799101.1, NW_015090839.1, NW_014006077.1, NC_046656.1, NC_041043.1, NC_040021.1, NC_045563.1, NC_044005.1, NW_006288624.1 ,NW_006501064.1,NW_010112985.1,NC_024307.1,NW_006209531.1,NW_018030485.1,NC_047090.1,NC_050191.1,NW_023365123.1,NW_015505036.1,NC_042571.1,NC_041720.1,NW_020448008.1,NW_006727797. 1、NW_012116265.1、NC_043979.1、NW_012150840.1、NW_013185668.1、NC_045964.1、NC_045701.1、NC_041066.1、NW_009270392.1、NW_007729665.1、NW_021604585.1、NC_044003.1、NC_045439.1、NC_046335.1、NW_ 008348666.1、NC_045481.1、NW_006921675.1、NW_020848012.1、NC_044039.1、NW_015553274.1、NW_019715876.1、NW_020172672.1、NC_044325.1、NW_019525097.1、NW_007014397.1、NC_046358.1、NW_020664447.1、NW_016107905.1, NC_047530.1, NW_004454173.1, NW_004443946.1, NW_019154104.1 and other NTCPs and variants thereof. In some embodiments, the NTCPs that are simultaneously combined with the delivery method and the vector are human mammalian and avian NTCPs. In some preferred embodiments, the NTCPs that are simultaneously combined with the delivery method and the vector are human NTCP (an exemplary amino acid sequence can be found in SEQ ID NO: 7, NCBI Reference Sequence: NM_003049), chimpanzee NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_510035), tree shrew NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006171503), and NTCPs of bonobos, tree shrews, marmots, arctic ground shrews, ground shrews, bats, woolly monkeys, capuchin monkeys, squirrel monkeys, and ducks. In some further preferred embodiments, the NTCP that is simultaneously combined with the delivery method and the vector is human NTCP (an exemplary amino acid sequence can be found in SEQ ID NO: 7, NCBI Reference Sequence: NM_003049), chimpanzee NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_510035), or tree shrew NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: XM_006171503). In some more preferred embodiments, the NTCP that is simultaneously combined with the delivery method and the vector is human NTCP (an exemplary amino acid sequence can be found in NCBI Reference Sequence: NM_003049).

[0159] The OATP-targeted delivery vectors include, but are not limited to, polypeptides, small molecule drugs, proteins, antibodies, nucleotides, and other forms of carriers, characterized by being able to bind to OATP and deliver the carried drug or group into cells and / or organs. Preferably, the delivery vectors can also target or bind to NTCP. The polypeptides, small molecule drugs, proteins, antibodies, nucleotides, and other forms of OATP-targeted delivery vectors and variants thereof at least retain the biological activity of binding to OATP and optionally also targeting or binding to NTCP to deliver the carried drug or group into cells, and / or organs, and / or systems. In some preferred embodiments herein, the polypeptides, small molecule drugs, proteins, antibodies, nucleotides, and other forms of OATP-targeted delivery vectors and variants thereof at least retain the biological activity of binding to OATP1B3, and / or OATP1B1, and / or OATP1B2, and delivering the carried drug or group into hepatocytes, and / or the liver, and / or the digestive system. In some more preferred embodiments herein, the polypeptides, small molecule drugs, proteins, antibodies, nucleotides and other forms of OATP-targeted delivery vectors and variants thereof at least retain the biological activity of binding to human OATP1B3 and / or human OATP1B1 and delivering the carried drugs or groups into human hepatocytes and / or human liver and / or human digestive system. In some embodiments herein, the OATP-targeted delivery vectors include but are not limited to polypeptides, antibodies, proteins, small molecule drugs, nucleotides and other types of compounds or molecules obtained through design and / or screening that can bind to or interact with OATP, currently known agents that bind to or interact with OATP, and polypeptides derived from the surface antigen of hepadnavirus.

[0160] In some embodiments herein, the OATP targeted delivery vectors described herein can be prepared by designing and / or screening polypeptides, antibodies, proteins, small molecule drugs, nucleotides, and other types of compounds or molecules that can bind to or interact with OATP and optionally bind to or interact with NTCP. In some preferred embodiments herein, the OATP targeted delivery vectors described herein can be prepared by designing and / or screening polypeptides, antibodies, proteins, small molecule drugs, nucleotides, and other types of compounds or molecules that can bind to or interact with OATP1B1 and / or OATP1B3. In some embodiments herein, polypeptides that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by designing and / or screening include, but are not limited to, polypeptides or polypeptide modifications obtained by targeting OATP1B1 and / or OATP1B3, including, but not limited to, random peptide libraries, combinatorial peptide libraries, synthetic peptide libraries, and other peptide library technologies, phage display technologies, yeast display technologies, viral display technologies, and other peptide screening technologies. In some embodiments herein, antibodies that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by design and / or screening include, but are not limited to, anti-OATP1B1 and / or anti-OATP1B3 antibody serum, polyclonal antibodies, monoclonal antibodies, antibody fragments, or antibody modifications obtained by immunizing animals with OATP1B1 and / or OATP1B3 proteins, protein fragments, or fusion proteins, or polyclonal antibodies, monoclonal antibodies, humanized antibodies, fully humanized antibodies, and other antibodies, or antibody fragments, or antibody modifications obtained by hybridoma technology, phage display technology, yeast display technology, immune library technology, ribosome display technology, mRNA display screening technology, Nanobody (Nb) screening method, transgenic mouse fully human antibody screening technology, single-cell photoconductive technology, flow cytometry technology, and other antibody screening technologies. In some embodiments herein, proteins that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by design and / or screening include, but are not limited to, proteins, amino acid fragments, or protein modifications obtained by targeting OATP1B1 and / or OATP1B3 through phage display technology, yeast display technology, cell display technology, random library, and other protein-protein interaction screening technologies. In some embodiments herein, nucleotides that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by design and / or screening include, but are not limited to, polyDNA, oligoDNA, polyRNA, oligoRNA, deoxyribonucleic acid-protein complexes (DNPs), or nucleic acid modifications obtained by screening using OATP1B1 and / or OATP1B3 as targets.In some embodiments herein, small molecules that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by design and / or screening include but are not limited to small molecules that can bind to or interact with OATP1B1 and / or OATP1B3 obtained by design or compound screening. In some preferred embodiments, polypeptides, antibodies, proteins, small molecule drugs, nucleotides and other types of compounds or molecules that can bind to or interact with human OATP1B1 and / or human OATP1B3 obtained by including but not limited to the above-mentioned design and / or screening techniques are used as delivery vectors targeting OATP. In some embodiments herein, the OATP-targeted delivery vectors described herein can be prepared by utilizing or modifying currently known agents that bind to or interact with OATP. In some preferred embodiments herein, the OATP-targeted delivery vectors described herein can be prepared by utilizing or modifying currently known agents that bind to or interact with OATP1B1 and / or OATP1B3. Agents currently known to interact with OATP1B1 and / or OATP1B3 include, but are not limited to, statins (including but not limited to atorvastatin, pitavastatin, rosuvastatin, simvastatin, pravastatin, and other statins), sartans (including but not limited to valsartan, olmesartan, telmisartan, and other sartans), bosentan, docetaxel, fexofenadine, glecaprevir, glyburide (glibenclamide), grazoprevir, nateglinide, paclitaxel, paritaprevir, repaglinide, simeprevir, voxilaprevir, steroid hormone metabolites, thyroid hormone metabolites, inflammatory mediators, bile acids, bilirubin, bromosulfophthalein, estradiol-17β-glucuronide, cholecystokinin octapeptide, estradiol-3-sulfate, liver function markers, mushroom toxins, antibiotics, antiviral drugs, anticancer drugs, Myrcludex B (bulevirtide), and other agents known to interact with OATP1B1 and / or OATP1B3.The aforementioned agents can be tested for their ability to bind to OATP1B1 and / or OATP1B3 and deliver drugs, or they can be modified to possess the ability to bind to OATP1B1 and / or OATP1B3 and deliver drugs. In some more preferred embodiments herein, the OATP-targeted delivery vectors described herein can be prepared by utilizing or modifying currently known agents that bind to or interact with human OATP1B1 and / or human OATP1B3.

[0161] The OATP-targeted delivery vector contains drugs or groups, including but not limited to small molecule drugs, nucleotides, proteins, antibodies, polypeptides, or other drugs or groups, characterized in that the drugs or groups can be linked to the vector via covalent and / or non-covalent bonds. The drugs or groups are linked to the vector, and through the combination or interaction between the vector and the OATP, the drugs or groups linked to the vector are delivered into cells, organs, or tissues. The drugs or groups can be linked to the OATP-targeted delivery vector via, but not limited to, covalent and / or non-covalent bonds or other means. The covalent bonds include but are not limited to peptide bonds, ester bonds, amide bonds, disulfide bonds, polar bonds, non-polar bonds, coordination bonds, single bonds, double bonds, triple bonds, σ bonds, π bonds, and other covalent bonds. The non-covalent bonds include but are not limited to hydrogen bonds, ion-dipole interactions, dipole-dipole interactions, charge interactions, hydrophobic interactions, hydrophilic interactions, van der Waals forces, ionic bonds, salt bonds, intermolecular forces, protein interactions, protein-nucleic acid interactions, antigen-antibody interactions, receptor-ligand interactions, and other non-covalent interactions. The OATP targeted delivery vector and the drug or group can be directly linked or linked through a linker. The small molecule drug or group includes but is not limited to fluorescent groups, toxins, chemotherapeutic drugs, signal regulatory molecules, cell signal agonists, cell signal inhibitors and other small molecule drugs or groups. The nucleotides include but are not limited to DNA, RNA and other nucleotide molecules or variants, including but not limited to oligo-DNA, oligo-RNA, poly-DNA, poly-RNA and other forms of DNA and / or RNA molecules. These nucleotide molecules have the functions of changing and regulating gene sequences, gene editing, gene transcription, mRNA translation, and their functions include but are not limited to transferring exogenous genes, protein coding, up-regulating or down-regulating target gene expression. The nucleic acid molecules include plasmids, DNA gene fragments, RNA gene fragments, antisense nucleic acids, ribozymes, RNA interference, gene editing vectors and other forms of nucleic acid molecules. In some more preferred embodiments herein, the nucleotides carried by the vector are protein-coding DNA or RNA, antisense nucleic acids, nucleic acids for RNA interference, and nucleotides for gene editing. In some embodiments herein, the nucleic acids carried by the vector also include but are not limited to viruses. The OATP targeting vector can be linked to a virus, or the vector can be fused with a viral protein for expression, or the gene encoding the OATP targeting vector can be inserted into the virus, or the vector can be linked to the virus in other ways, so that the virus acquires the biological activity of infecting cells expressing OATP, thereby delivering the nucleic acid code or fragment carried by the virus into cells, organs, or tissues. The virus includes, but is not limited to, adenovirus, lentivirus, retrovirus, adeno-associated virus, poxvirus, stomatitis virus, herpes virus, rabies virus vector, and other viruses.The proteins include, but are not limited to, proteins that can regulate cellular physiological functions, proteins with pharmacological effects, proteins that can kill target cells, and proteins with other activities. The antibodies include, but are not limited to, antibodies that can regulate cellular physiological functions after binding to the target antigen, antibodies with pharmacological effects, antibodies that can kill target cells, antibodies that regulate immunity, and antibodies with other activities. The antibodies include, but are not limited to, polyclonal antibodies, monoclonal antibodies, bispecific antibodies, trispecific antibodies, tetraspecific antibodies, humanized antibodies, fully humanized antibodies, antibody fragments, and other antibodies. These proteins or antibodies and their fragments can be expressed by (but not limited to) fusion with the vector, or coupled to the vector after expression, or coupled to the vector by other means.

[0162] The OATP targeted delivery method and carrier delivery drug can be used for mammals, birds, fish, amphibians and other animals. The mammals include but are not limited to primates, tree shrews, rodents, bats, artiodactyla, carnivora and other mammals. The birds include but are not limited to ducks, herons and other avian animals. The primates include but are not limited to hominoid species, New World monkeys, Old World monkeys and other primates. The hominoid species include but are not limited to humans, chimpanzees, gorillas, gibbons, orangutans and other hominoid species. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora. In some preferred embodiments herein, the vector is used for drug delivery in humans, chimpanzees, bonobos, tree shrews, ducks, marmots, Arctic shrews, shrews, bats, woolly monkeys, capuchin monkeys, and squirrel monkeys.In some more preferred embodiments herein, the vector is used for drug delivery in humans.

[0163] In the OATP-targeted delivery method and vector, the systems into which the drug is delivered include but are not limited to the digestive system, respiratory system, circulatory system, nervous system, musculoskeletal system, urinary system, reproductive system, endocrine system, and other systems; the organs and tissues into which the drug is delivered include but are not limited to the liver, kidney, lung, heart, muscle, fat, brain, nerves, intestines, pancreas, and other organs or tissues. The delivered drug preferably enters the digestive system, and the delivered drug preferably enters the liver. In some embodiments herein, the systems into which the drug is delivered using the vector include but are not limited to the digestive system, respiratory system, circulatory system, nervous system, musculoskeletal system, urinary system, reproductive system, endocrine system, and other systems. In some embodiments herein, the organs and tissues into which the drug is delivered using the vector include but are not limited to the liver, kidney, lung, heart, muscle, fat, brain, nerves, intestines, pancreas, and other organs or tissues. In some embodiments herein, the cells into which the drug is delivered using the vector include but are not limited to hepatocytes, lung cells, kidney cells, nerve cells, muscle cells, adipocytes, blood cells, lymphocytes, and other cells. In some preferred embodiments herein, the vector is used to deliver the drug into the digestive system, and / or liver, and / or hepatocytes.

[0164] In a preferred embodiment, the OATP-targeted drug delivery vector described herein is preferably an OATP-targeted delivery polypeptide vector derived from the surface antigen of hepadnavirus as described below.

[0165] OATP-targeted delivery peptide vector derived from hepadnavirus surface antigen

[0166] In some embodiments herein, the OATP targeted delivery vector provided herein includes but is not limited to a polypeptide derived from the surface antigen of hepadnavirus, which polypeptide at least retains the biological activity of binding to OATP and delivering the carried drug or group into cells, that is, an OATP targeted delivery polypeptide vector derived from the surface antigen of hepadnavirus, referred to as OATP targeted delivery polypeptide vector.

[0167] The hepadnavirus includes but is not limited to Orthohepadnavirus, Avihepadnavirus, Metahepadnavirus, Herpetohepadnavirus and other hepadnaviruses. The orthohepadnavirus includes but is not limited to orthohepadnaviruses that infect humans (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkey, woodchuck hepatitis virus (WHV), Arctic ground shrew, ground shrew (Ground squirrel hepatitis virus, GSHV), tree shrew, chimpanzee, bat (BHV) and other animals. The genome sequence can be found in, but is not limited to, GenBank accession numbers AF046996 (WMHBV), NC_043528 (CMHBV), NC_004107 (WHV), NC_001484 (GSHV), and JX941468 (BHV). The Avihepadnavirus includes but is not limited to Avihepadnavirus that infects ducks (DHBV), herons (HHBV) and other birds, and exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers NC_001344 (DHBV) and NC_001486 (HHBV). The human-infecting proto-hepadnavirus (HBV) includes but is not limited to genotypes A, B, C, D, E, F, G, H and I genotype HBV, and exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G) and AB179747 (genotype H).

[0168] The amino acid sequence of the OATP targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus is derived from the viral envelope protein (envelope protein), also known as the surface antigen (surface antigen) or surface protein (surface protein). The hepadnavirus includes but is not limited to humans (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkey, woodchuck hepatitis virus (WHV), Arctic ground shrew, ground shrew (Ground squirrel hepatitis virus, GSHV), tree shrew, chimpanzee, bat (BHV) and other animals' original hepadnavirus, as well as duck HBV (DHBV), heron HBV (HHBV) and other birds' Avihepadnavirus. In some preferred embodiments herein, the amino acid sequence of the OATP targeted delivery polypeptide vector is derived from the amino acid sequence of the viral surface antigen Pre-S region, including but not limited to the exemplary surface antigen Pre-S amino acid sequence SEQ ID NO: 8-22 and variants thereof. In some preferred embodiments herein, the OATP targeted delivery polypeptide vector is a polypeptide vector derived from the Pre-S1 region of the human HBV surface antigen, including but not limited to the amino acid sequence of the genotype A, B, C, D, E, F, G, H and I genotype HBVPre-S1 region and variants thereof, including but not limited to the exemplary HBV surface antigen Pre-S1 amino acid sequence SEQ ID NO: 15-22 and variants thereof. In some embodiments herein, the OATP targeted delivery polypeptide vector may be derived from the pre-S1 region of the surface antigen of HBV genotype C. In some embodiments, the OATP targeted delivery polypeptide vector may consist of 10-118 amino acids in length. For example, the length of the polypeptide may be 15-100, 15-80, 20-100, 20-80, 20-60, 25-60, 30-60, 35-60 or 40-60 (including all integers between these ranges) amino acids. In some embodiments, the length of the OATP targeted delivery polypeptide vector may be at least 20 amino acids, for example, at least 25, 30, 35 or 40 amino acids. In some embodiments, the length of the OATP targeted delivery polypeptide vector may be 20, 25, 30, 35, 40, 47, 55 or 60 amino acids. In some embodiments, the length of the OATP targeted delivery polypeptide vector may be 47 amino acids.Variants of the OATP targeted delivery polypeptide vector with different lengths retain one or more biological activities associated with the corresponding polypeptide, including at least the biological activities of binding to OATP (particularly OATP1B1 and OATP1B3, especially human OATP1B1 and human OATP1B3) and transporting drugs into cells.

[0169] The terms "OATP targeted delivery polypeptide vector derived from the surface antigen of hepadnavirus", "polypeptide vector derived from the surface antigen of hepadnavirus", and "polypeptide vector derived from hepadnavirus" mean that the origin or source of the polypeptide vector is hepadnavirus, including but not limited to natural, recombinant, synthetic or purified polypeptide vectors, including but not limited to full-length natural HBV polypeptides or fragments thereof, and variants of the full-length natural polypeptides or fragments thereof. In some embodiments, the polypeptide vector derived from the hepadnavirus can be derived from any hepadnavirus, including but not limited to the orthohepadnavirus (OHV) that infects mammals, the Avihepadnavirus (AHV) that infects birds, the Metahepadnavirus that infects fish, and the Herpetohepadnavirus that infects amphibians; in some embodiments, the polypeptide vector derived from the hepadnavirus can be derived from any hepadnavirus surface antigen; in some embodiments, the polypeptide vector derived from the hepadnavirus can be derived from any orthohepadnavirus surface antigen; in some embodiments, the polypeptide vector derived from the hepadnavirus can be derived from any human hepatitis B virus (HBV) surface antigen, including but not limited to HBV surface antigens derived from genotypes A, B, C, D, E, F, G, H and I; in some embodiments, the polypeptide vector derived from the hepadnavirus can be derived from the Pre-S1 region of any human hepatitis B virus (HBV) surface antigen, that is, an OATP targeted delivery polypeptide vector derived from HBVPre-S1.

[0170] The term "HBVPre-S1-derived OATP targeted delivery polypeptide vector" refers to a polypeptide whose origin or source is the HBV surface antigen Pre-S1 region, including but not limited to natural, recombinant, synthetic or purified polypeptides, including but not limited to the full-length natural HBV surface antigen Pre-S1 region or fragments thereof and variants thereof, full-length non-natural HBV surface antigen Pre-S1 region or fragments thereof and variants thereof, non-full-length natural HBV surface antigen Pre-S1 region or fragments thereof and variants thereof, non-full-length non-natural HBV surface antigen Pre-S1 region or fragments thereof and variants thereof, and other forms of variants. In some embodiments, the HBVPre-S1-derived OATP targeted delivery polypeptide vector may contain the entire surface antigen pre-S1 region of any HBV genotype surface antigen pre-S1 region starting from the second amino acid G (glycine). In certain embodiments, the HBVPre-S1-derived OATP targeted delivery polypeptide vector may be derived from the pre-S1 region of any one of the surface antigens of HBV genotypes A, B, C, D, E, F, G, H and I. Exemplary genomic sequences of these HBV genotypes can be found, but are not limited to, GenBank accession numbers KC875260, AY220704, AF461363, AY796030, AB205129, DQ823095, HE981176, and AB179747. Exemplary amino acid sequences of the pre-S regions of their surface antigens can be found, but are not limited to, SEQ ID NOs: 8-22 and variants thereof. In certain embodiments, the OATP-targeted delivery polypeptide vector derived from HBVPre-S1 can be derived from the pre-S1 region of the HBV surface antigen of genotype C. The OATP-targeted delivery polypeptide vector derived from HBVPre-S1 at least retains the biological activity of binding to OATPs, particularly retains the biological activity of binding to OATP1B1, and / or OATP1B2, and / or OATP1B3, and particularly retains the biological activity of binding to human OATP1B1 and / or human OATP1B3, and can transport the carried drug or group into cells.

[0171] In some embodiments herein, the OATP targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus has a structure shown in the following formula (I):

[0172] Wherein G is glycine; the core sequence is an amino acid sequence derived from the surface antigen of a hepadnavirus, used as a carrier; X is an amino acid residue having two amino groups or two carboxyl groups or other groups available for linking reactions; R1 is a modification of the N-terminus of the carrier; R2 is absent or is a flanking sequence or other group; and R3 is a carried pharmacophore or a linking group that can interact with a drug or group.

[0173] The core amino acid sequence is derived from the envelope protein of the hepadnavirus, also known as the surface antigen or surface protein. The hepadnaviruses include, but are not limited to, human (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkey, woodchuck hepatitis virus (WHV), Arctic ground shrew, ground shrew (GSHV), tree shrew, chimpanzee, bat hepatitis virus (BHV) and other animals, as well as Avihepadnavirus of duck (DHBV), heron (HHBV) and other birds. In some preferred embodiments herein, the polypeptide vector derived from the hepadnaviridae has a core amino acid sequence derived from the amino acid sequence of the viral surface antigen Pre-S region, including but not limited to the exemplary surface antigen Pre-S amino acid sequence SEQ ID NO: 8-22 and variants thereof. In some preferred embodiments herein, the polypeptide vector core sequence is derived from a polypeptide of the human HBV surface antigen Pre-S region amino acid sequence, including but not limited to genotype A, B, C, D, E, F, G, H and I genotype HBVPre-S1 region amino acid sequence, including but not limited to the exemplary HBV surface antigen Pre-S amino acid sequence SEQ ID NO: 15-22 and variants thereof. In some embodiments herein, the polypeptide vector core sequence is derived from a human genotype C HBV polypeptide comprising a surface antigen pre-S1 region amino acid sequence, including but not limited to the exemplary amino acid sequence SEQ ID NO: 17 and variants thereof. In some embodiments herein, the polypeptide vector core sequence is derived from a human genotype C HBV polypeptide comprising amino acids 3-119, 4-119, 3-69, 4-69, 3-59, 4-59, 3-48, 4-48, 14-119, 14-88, 14-72, 14-67, 14-59, 14-56, 14-55, 14-53, 14-48, 14-47, 14-45, 14-42, 14-37, and 14-32 of the surface antigen pre-S1 region, exemplary sequences of which include, but are not limited to, SEQ ID NOs: 81-102 and variants thereof.In some embodiments, the vector core sequence comprises at least amino acid residues 14-32 of the N-terminal pre-S1 region of the genotype C HBV surface antigen described herein. In preferred embodiments, the core sequence may be 19-106 amino acid residues in length. In particularly preferred embodiments, the core sequence is, for example, but not limited to, the sequence exemplified in SEQ ID NO: 97, which comprises at least, but not limited to, amino acid residues 1-19 of the sequence exemplified in SEQ ID NO: 97. Preferably, the core sequence comprises at least, but not limited to, amino acid residues 1 to 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 of the sequence exemplified in SEQ ID NO: 30. In some embodiments, the core sequence comprises a fragment of at least amino acid residues 14-32 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I, a fragment of at least amino acid residues 3-21 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and a fragment of at least amino acid residues 13-31 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E and G. Further preferably, the core sequence comprises at least amino acid residues 14-48 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I, at least amino acid residues 3-37 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and at least amino acid residues 13-47 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E and G. Exemplary sequences thereof include, but are not limited to, SEQ ID NOs: 103-110.

[0174] As used herein, a "variant" of a core sequence refers to a polypeptide that differs in amino acid sequence from a given polypeptide but retains the biological activity of binding to an OATP and transporting a drug or moiety into cells, particularly the biological activity of binding to OATP1B3, and / or OATP1B2, and / or OATP1B1 and transporting a drug or moiety into hepatocytes, and particularly the biological activity of binding to human OATP1B3 and / or OATP1B1 and transporting a drug or moiety into human hepatocytes. For example, in some embodiments, amino acid substitutions or mutation combinations of N15D, F25L, G35K, N39E, N46K, F45L, N48H, N48Y, N48K, D50A, H51Q, H51N, E54K, E54D, A55S, N56K, N56D, and Q57K can be introduced into the core sequence derived from the amino acid sequence 14-59 of the HBVPre-S1 region of genotype C. Example sequences thereof include, but are not limited to, SEQ ID NOs: 111-128. The term "variant" also includes homologous core sequences found in different viral species, strains, or subtypes of the genus Hepadnavirus. HBV is divided into four major serotypes (adr, adw, ayr, and ayw) based on antigenic epitopes on its envelope protein, and into nine genotypes (AI) based on the variability of the entire nucleotide sequence in the genome. Therefore, the term "variant" includes any of these homologous core sequences found in HBV subtypes. "Variant" may also include core sequences and variants thereof with naturally occurring flanking amino acid sequences from any of these HBV subtypes added to the N- and / or C-termini. For example, in some embodiments, a polypeptide derived from amino acid sequences 13-48 of the pre-S1 region of the genotype C surface antigen can be fused with a fragment of at least amino acid residues 3-12 of the pre-S1 region of the genotype A, B, F, H, or I HBV surface antigen, or a fragment of at least amino acid residues 3-11 of the pre-S1 region of the genotype E or G HBV surface antigen to form a fusion core sequence. Example sequences include, but are not limited to, SEQ ID NOs: 129-134. In some embodiments, the core sequence described herein may comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 81-128 and a naturally occurring amino acid sequence flanking the N- and / or C-terminus of a surface antigen derived from any of HBV genotypes A to I. In some embodiments, the naturally occurring flanking amino acid sequence may be derived from, but is not limited to, an HBV surface antigen amino acid sequence having GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G), or AB179747 (genotype H).For example, the core sequence described herein may comprise the amino acid sequence set forth in SEQ ID NO: 97 and, at its N- and / or C-terminus, contain naturally occurring flanking amino acid sequences from the pre-S1 region of HBV genotype C. Alternatively, the core sequence described herein may comprise the amino acid sequence set forth in SEQ ID NO: 97 and, at its N- and / or C-terminus, contain naturally occurring flanking amino acid sequences from the pre-S1 region of any of HBV genotypes A, B, C, D, E, F, G, and H. In some embodiments, the N- and / or C-terminus of the core sequence described herein may independently contain naturally occurring flanking amino acid sequences of 1-10 amino acids in length, such as 1-8, 1-5, or 1-3 amino acids in length (including all integers within these ranges). For example, the core sequence described herein may comprise the amino acid sequence set forth in SEQ ID NO: 97 and, at its N-terminus, contain naturally occurring flanking amino acid sequences of 10 amino acids in length from the pre-S1 region of HBV genotype C. In other words, the core sequence may comprise amino acids 3-48 (SEQ ID NO: 87) of the pre-S1 region of HBV genotype C. As another example, the core sequence described herein may comprise the amino acid sequence set forth in SEQ ID NO: 97 and, at its N-terminus, a naturally occurring flanking amino acid sequence of 9 amino acids from the pre-S1 region of HBV genotypes E or G. In other words, the polypeptide may comprise amino acids 14-48 of the pre-S1 region of HBV genotype C and amino acids 3-12 of the pre-S1 region of HBV genotypes E or G (SEQ ID NOs: 131 and 133). It should be understood that any of the vector core sequences described herein may have naturally occurring flanking amino acid sequences of any length extending from its N- and / or C-terminus, and that the resulting core sequence retains the biological activity of the original polypeptide vector, including at least binding to OATPs (particularly OATP1B1 and OATP1B3) and transporting drugs or groups into cells. "Variants" of the core sequence also include those having one or more amino acid deletions, substitutions, or insertions, while retaining the multiple biological activities of the original polypeptide vector, including at least binding to OATPs (particularly OATP1B1 and OATP1B3, especially human OATP1B1 and human OATP1B3) and transporting drugs or groups into cells. For example, in some embodiments, relative to the core sequence of the pre-S1 region of HBV, the polypeptides described herein may have 1-30, e.g., 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions.For example, in some embodiments, the polypeptides described herein may have 1-30, e.g., 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the core sequence of any one of SEQ ID NOs: 81-128. For example, in some embodiments, the vector core sequence described herein may have 1-30, e.g., 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the amino acid sequence of SEQ ID NO: 166. For example, in some embodiments, the vector core sequence described herein may have 1-3 amino acid deletions, substitutions, or insertions relative to the amino acid sequence of SEQ ID NO: 97. For example, in certain embodiments, the vector core sequence described herein has 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions or insertions at the N-terminus and / or C-terminus relative to the amino acid sequence selected from any one of SEQ ID NOs: 81-128. For example, the vector core sequence described herein may contain an amino acid sequence selected from any one of SEQ ID NOs: 81-102. In various embodiments, the vector core sequence described herein may have at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any one of the vector core sequences described herein. For example, the polypeptide may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any of the exemplary sequences in SEQ ID NOs: 81-134. In some embodiments, the vector core sequence may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any of the exemplary sequences in SEQ ID NOs: 81-134. In some embodiments, the vector core sequence may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence exemplified in SEQ ID NO: 97. Variants having a certain degree of sequence identity with the core sequence described herein allow the resulting polypeptide vector to at least retain the biological activity of binding to OATPs (particularly OATP1B1 and OATP1B3, especially human OATP1B1 and human OATP1B3) and transporting drugs or groups into cells.

[0175] R1 is a required N-terminal modification group for OATP-targeted delivery polypeptide vectors derived from the surface antigen of the hepadnavirus. In some embodiments, R1 may be modified with a hydrophobic group, such as, but not limited to, myristic acid (myr), palmitic acid (plam), stearic acid (stearoyl), cholesterol (cho1), oleic acid, linoleic acid, polyethylene glycol (PEG), arachidonic acid, or other hydrophobic groups. In some embodiments, the hydrophobic group R1 may be selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In some embodiments, the hydrophobic group R1 may be myristic acid. In certain embodiments, the core sequence of the vector described herein comprises an amino acid sequence selected from any one of SEQ ID NOs: 81-134, wherein the N-terminal G of the polypeptide may be modified with a hydrophobic group selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In certain embodiments, the core sequence of the polypeptide vector described herein may comprise an amino acid sequence selected from any one of SEQ ID NOs: 81-134, wherein the N-terminal G of the polypeptide vector may be modified with a hydrophobic group selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In certain embodiments, the core sequence of the polypeptide vector described herein may comprise an amino acid sequence selected from any one of SEQ ID NOs: 81-134, wherein the N-terminal G of the polypeptide vector may be modified with myristoylation. In some embodiments, the core sequence of the polypeptide vector described herein may contain the amino acid sequence shown in SEQ ID NO: 97 or SEQ ID NO: 94, wherein the N-terminal G of the polypeptide vector may be myristoylated.

[0176] In formula (I), G is connected to the core sequence via an amide bond; X is connected to the core sequence via an amide bond.

[0177] R2 is a flanking sequence or other group, and R2 can be deleted. In some embodiments herein, R2 is derived from an amino acid sequence derived from a surface antigen of a hepadnavirus, including but not limited to humans (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkey, woodchuck hepatitis virus (WHV), Arctic ground shrew, ground shrew (Ground squirrel hepatitis virus, GSHV), tree shrew, chimpanzee, bat (BHV) and other animals' original hepadnavirus, and duck HBV (DHBV), heron HBV (HHBV) and other birds' Avihepadnavirus. In some preferred embodiments herein, in the polypeptide vector derived from the hepadnaviridae, its R2 flanking amino acid sequence is derived from the amino acid sequence of the viral surface antigen Pre-S region, including but not limited to the exemplary surface antigen Pre-S amino acid sequence SEQ ID NO: 8-22 and variants thereof. In some preferred embodiments herein, the R2 flanking sequence of the polypeptide vector is a polypeptide derived from the amino acid sequence of the human HBV surface antigen Pre-S region, including but not limited to genotype A, B, C, D, E, F, G, H and I genotype HBVP Pre-S1 region amino acid sequence or a fragment thereof, including but not limited to the exemplary HBV surface antigen Pre-S amino acid sequence SEQ ID NO: 15-22 and variants thereof. In some embodiments herein, the polypeptide vector R2 flanking sequence is derived from a human genotype C HBV polypeptide comprising a surface antigen pre-S1 region amino acid sequence, including but not limited to the exemplary amino acid sequence SEQ ID NO: 17 and variants thereof. In some embodiments herein, the polypeptide carrier R2 flanking sequence is derived from a human genotype C HBV polypeptide comprising amino acids 50-119, 50-88, 50-72, 50-69, 50-67, 58-59, 50-59, or 50-52 of the pre-S1 region of the surface antigen, exemplary sequences of which include, but are not limited to, SEQ ID NOs: 135-142 and variants thereof. In some embodiments herein, the R2 flanking sequence contains no amino acids or contains amino acid residues 50-52 of the N-terminal pre-S1 region of the genotype C HBV surface antigen as described herein. In some embodiments herein, the R2 flanking sequence may be 0-70 amino acid residues in length.In some preferred embodiments, the R2 flanking sequence contains at least amino acid residues 1-3 of SEQ ID NO: 141, preferably at least amino acid residues 1 to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of SEQ ID NO: 141; in some embodiments, the R2 flanking sequence is as shown in SEQ ID NO: 141. In some embodiments, the fragment comprises at least amino acid residues 50-52 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I as described herein, at least amino acid residues 39-41 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and at least amino acid residues 49-51 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E and G. Further preferably, the fragment contains a fragment of at least amino acid residues 50-59 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype A, B, F, H, and I, a fragment of at least amino acid residues 39-48 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and a fragment of at least amino acid residues 49-58 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype E and G, and example sequences thereof include but are not limited to SEQ ID NOs: 143-149.

[0178] As used herein, a "variant" related to the R2 flanking sequence described herein, the flanking sequence derived from HBV, or the HBV-derived flanking sequence refers to a variant that differs in amino acid sequence from a given flanking sequence (i.e., the flanking sequence described herein, the flanking sequence derived from HBV, or the HBV-derived flanking sequence), but retains the biological activity of binding to OATPs and transporting drugs or groups into cells, particularly the biological activity of binding to OATP1B3, and / or OATP1B2, and / or OATP1B1, and transporting drugs or groups into hepatocytes, and particularly the biological activity of binding to human OATP1B3 and / or OATP1B1, and transporting drugs or groups into human hepatocytes. For example, in some embodiments, amino acid substitutions or mutations such as D50A, H51Q or H51N, E54K or E54D, A55S, N56K or N56D, or Q57K can be introduced into a polypeptide derived from the amino acid sequence of positions 50-59 of the HBVPre-S1 region of genotype C. Example sequences include, but are not limited to, SEQ ID NOs: 150-158. The term "variant" also includes homologous core sequences found in different virus species, strains or subtypes of the genus Hepadnavirus. Based on the antigenic epitopes on its envelope protein, HBV is divided into four major serotypes (adr, adw, ayr and ayw), and based on the variability of the entire nucleotide sequence in the genome, HBV is divided into 9 genotypes (AI). Therefore, the term "variant" includes any of these homologous flanking sequences found in HBV subtypes. "Variant" may also include flanking sequences and variants thereof with natural flanking amino acid sequences from any of these HBV subtypes added to the N and / or C termini. In some embodiments, the natural flanking amino acid sequence may be derived from HBV surface antigen amino acid sequences including but not limited to GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G) or AB179747 (genotype H).

[0179] In some embodiments, the N- and / or C-termini of the flanking sequences described herein may independently contain a native flanking amino acid sequence of 1-10, such as 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acids in length. "Variants" of the flanking sequences also include those having one or more amino acid deletions, substitutions, or insertions, while the resulting flanking sequences retain the various biological activities of the original polypeptide vector, including at least binding to OATPs (particularly OATP1B1 and OATP1B3, especially human OATP1B1 and human OATP1B3) and transporting drugs or groups into cells. For example, in some embodiments, relative to a sequence from the pre-S1 region of HBV, the flanking sequences described herein may have 1-30, such as 1-20, 1-20, 1-10, 1-5, or 1 (including all integers within these ranges) amino acid deletions, substitutions, or insertions. For example, in some embodiments, the flanking sequences described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the amino acid sequence set forth in any one of SEQ ID NOs: 135-158. For example, in some embodiments, the vector core flanking sequences described herein may have 1-10, such as 1-10, 1-8, 1-5, 1-3, or 1 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the amino acid sequence of SEQ ID NO: 141. For example, in some embodiments, the vector core flanking sequences described herein may have 1-3 amino acid deletions, substitutions, or insertions relative to the amino acid sequence of SEQ ID NO: 141. For example, in certain embodiments, the vector core sequence described herein has 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions or insertions at the C-terminus relative to the amino acid sequence shown in any one of SEQ ID NOs: 135-158. For example, the vector core sequence described herein may contain an amino acid sequence shown in any one of SEQ ID NOs: 135-142.

[0180] In various embodiments, the vector flanking sequences described herein can be at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any of the flanking core sequences described herein. For example, the flanking sequence can comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 135-158. In some embodiments, the flanking core sequence may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any one of SEQ ID NOs: 135-158. In some embodiments, the flanking core sequence may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 141. Variants having certain sequence identities to the flanking sequences described herein allow the resulting polypeptide vector to at least retain the biological activity of binding to OATPs (particularly OATP1B1 and OATP1B3, particularly human OATP1B1 and human OATP1B3) and transporting drugs or groups into cells. In some embodiments, the C-terminus of the flanking sequence may be modified. The C-terminal modification of the flanking sequence may be selected from, but not limited to, amidation (amination), prenylation, and any other modification, or may be absent. For example, in some embodiments, the C-terminal modification of the flanking sequence is selected from amidation (NH2). In some embodiments herein, the flanking sequence is selected from SEQ ID NO: 141, 158, 140, and its C-terminus is modified by amidation. In some embodiments herein, R2 may not contain a flanking sequence and is only a modifying group, or may be absent. In some embodiments, R2 may be amidated, prenylated, or other groups.

[0181] In formula (I), the amino acid having two amino groups in X can be, for example, lysine, arginine, histidine, or asparagine; the amino acid having two carboxyl groups can be, for example, aspartic acid or glutamic acid; and the amino acid having other groups available for linkage reactions can be, for example, methionine. Therefore, in formula (I) herein, R2 and R3 can react with the two groups of X to form an amide bond or other chemical bond for covalent linkage. These reactions include, but are not limited to, acylation reactions, nucleophilic reactions, sulfonylation reactions, and salt formation reactions. Preferably, when R2 is a flanking sequence, R2 and X are linked via an amide bond.

[0182] R3 is the carried pharmacophore, or a linker (i.e., a linker) that can interact with a drug or group. The pharmacophore is directly linked to the carrier, or indirectly linked via the linker, and the carrier portion binds or interacts with the OATP, delivering the pharmacophore, drug, or group linked to the carrier portion into cells, organs, or tissues. The pharmacophore, drug, or group can be linked to the OATP-targeted delivery polypeptide vector derived from the hepadnavirus surface antigen via, but not limited to, covalent bonds and / or non-covalent bonds or other means. For example, in some embodiments, R3 is the carried drug group, and the pharmacophore is linked to the carrier via a covalent bond, with or without a linker between the pharmacophore and the carrier. The covalent bond includes, but is not limited to, a peptide bond, an ester bond, an amide bond, a disulfide bond, a polar bond, a non-polar bond, a coordinate bond, a single bond, a double bond, a triple bond, a sigma bond, a pi bond, and other covalent bonds. In some embodiments, the pharmacophore and the carrier are covalently linked via a linker, which may be selected from, but not limited to, amino acid sequences, nucleotide sequences, polyethylene glycol, and other linkers. In some embodiments, the linker may be an amino acid sequence, which may be selected from, but not limited to, flexible linkers rich in glycine G (Gly, G) and / or serine (Ser, S) or other amino acid sequence linkers. In some embodiments, R3 is a linking group that can interact with the drug or group, and the linking group is connected to the drug or group via a non-covalent bond. The non-covalent bond includes, but is not limited to, hydrogen bonds, ion-dipole interactions, dipole-dipole interactions, charge interactions, hydrophobic interactions, hydrophilic interactions, van der Waals forces, ionic bonds, salt bonds, intermolecular forces, protein interactions, protein nucleic acid interactions, antigen-antibody interactions, receptor-ligand interactions, and other non-covalent bond interactions. The OATP targeted delivery polypeptide carrier derived from the surface antigen of the hepadnavirus and the drug or group may be directly linked or linked via a linker.

[0183] The pharmacophores, drugs, or groups carried by the OATP-targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus include, but are not limited to, small molecule drugs, nucleotides, proteins, antibodies, polypeptides, or other drugs or groups. The small molecule drugs or groups include, but are not limited to, fluorescent groups, toxins, chemotherapeutic drugs, signal regulatory molecules, cell signaling agonists, cell signaling inhibitors, and other small molecule drugs or groups. The nucleotides include, but are not limited to, DNA, RNA, and other nucleotide molecules or variants, including, but not limited to, oligo-DNA, oligo-RNA, poly-DNA, poly-RNA, and other forms of DNA and / or RNA molecules. These nucleotide molecules have the ability to alter and regulate gene sequences, gene editing, gene transcription, and mRNA translation, and their functions include, but are not limited to, the introduction of exogenous genes, protein encoding, and up-regulation or down-regulation of target gene expression. The nucleic acid molecules include plasmids, DNA gene fragments, RNA gene fragments, antisense nucleic acids, ribozymes, RNA interference, gene editing vectors, and other forms of nucleic acid molecules. In some more preferred embodiments herein, the nucleotides carried by the vector are protein-coding DNA or RNA, antisense nucleic acids, nucleic acids for RNA interference, or nucleotides for gene editing. In some embodiments herein, the nucleic acid carried by the OATP-targeted delivery polypeptide vector derived from the surface antigen of a hepadnavirus also includes, but is not limited to, a virus. The OATP-targeted delivery polypeptide vector derived from the surface antigen of a hepadnavirus can be conjugated to a virus, or the vector can be fused with a viral protein for expression, or the vector can be conjugated to a virus by other means, allowing the virus to acquire the biological activity of infecting cells expressing the OATP, thereby delivering the nucleic acid encoding or fragment carried by the virus into cells, organs, or tissues. Such viruses include, but are not limited to, adenoviruses, lentiviruses, retroviruses, adeno-associated viruses, poxviruses, stomatitis viruses, herpes viruses, rabies virus vectors, and other viruses. Such proteins include, but are not limited to, proteins that can regulate cellular physiological functions, proteins with pharmacological effects, proteins that can kill target cells, and proteins with other activities. The antibodies include, but are not limited to, antibodies that can regulate cellular physiological functions after binding to the target antigen, antibodies with pharmacological effects, antibodies that can kill target cells, antibodies that regulate immunity, and antibodies with other activities. The antibodies include, but are not limited to, polyclonal antibodies, monoclonal antibodies, bispecific antibodies, trispecific antibodies, tetraspecific antibodies, humanized antibodies, fully humanized antibodies, antibody fragments, and other antibodies. These proteins or antibodies and their fragments can be expressed by (but not limited to) fusion with the vector, or coupled to the vector after expression, or coupled to the vector by other means.

[0184] In some embodiments herein, R3 is a fluorescent group, including but not limited to FITC, CY3, and CY5. In one embodiment herein, R3 is a FITC fluorescent group. Fluorescent groups such as FITC can undergo a nucleophilic reaction with the ε-amino group of K to form a thiourea linkage.

[0185] In some embodiments herein, R3 is a nucleotide, including but not limited to DNA, RNA and other nucleotide molecules, including but not limited to oligo-DNA, oligo-RNA, poly-DNA, poly-RNA and other forms of DNA and / or RNA molecules. These nucleotide molecules have the ability to change and regulate gene sequences, gene editing, gene transcription, and mRNA translation, and their function is to introduce exogenous genes and up-regulate or down-regulate target gene expression. The above-mentioned nucleic acid molecules include plasmids, DNA gene fragments, RNA gene fragments, antisense nucleic acids, ribozymes, RNA interference, gene editing vectors and other forms of nucleic acid molecules.

[0186] In some embodiments herein, R3 can be a polypeptide, protein, or antibody used to modulate the physiological function of target cells or kill target cells. The polypeptide, protein, or antibody group can be expressed by (but not limited to) fusion with a polypeptide vector, or coupled by chemical means after expression.

[0187] In some embodiments herein, the R3 group can be a virus, which is expressed by fusing the polypeptide vector with a viral protein, or by linking or coupling the polypeptide vector to the virus through chemical means or the like.

[0188] In some embodiments herein, R3 is a linking group that interacts with the carried drug, and the group can interact with the carried drug via a covalent bond or a non-covalent bond to achieve a connection between the drug and the OATP targeting carrier.

[0189] In some embodiments herein, the OATP targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus has a core sequence containing an amino acid sequence selected from any one of SEQ ID NOs: 81-134 and variants thereof; R1 can be selected from any hydrophobic group selected from myristic acid, palmitic acid, stearic acid, polyethylene glycol and cholesterol; the R2 flanking sequence contains an amino acid sequence selected from any one of SEQ ID NOs: 135-158 or a variant thereof, and its C-terminus can be amidated, or is only a modifying group, or can be deleted; R3 can be selected from any pharmacophore of a small molecule drug, nucleotide, polypeptide, protein or antibody, or a linking group that can interact with any drug of a small molecule drug, nucleotide, polypeptide, protein or antibody.

[0190] In some preferred embodiments herein, the core sequence of the OATP targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus contains an amino acid sequence selected from any one of SEQ ID NOs: 97, 94, 103-128 and variants thereof; R1 can be selected from a myristic acid hydrophobic group; the R2 flanking sequence contains an amino acid sequence selected from any one of SEQ ID NOs: 141, 140, 143-158 and variants thereof, the C-terminus of which is amidated, or is only a modifying group, or can be deleted; R3 can be selected from any pharmacophore of a small molecule drug, nucleotide, polypeptide, protein or antibody, or a linking group that can interact with any drug of a small molecule drug, nucleotide, polypeptide, protein or antibody.

[0191] In some preferred embodiments herein, the core sequence of the OATP targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus contains an amino acid sequence selected from any one of SEQ ID NOs: 97 and 94 and variants thereof; R1 can be selected from a myristic acid hydrophobic group; the R2 flanking sequence contains an amino acid sequence selected from any one of SEQ ID NOs: 141 and 140 and variants thereof, the C-terminus of which is amidated, or is only a modifying group, or can be deleted; R3 can be selected from any pharmacophore of a small molecule drug, a nucleotide, a polypeptide, a protein or an antibody, or a linking group that can act with any drug of a small molecule drug, a nucleotide, a polypeptide, a protein or an antibody.

[0192] In some preferred embodiments herein, the core sequence of the OATP-targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus comprises an amino acid sequence selected from the exemplary sequence set forth in SEQ ID NO: 97 and variants thereof; R1 is selected from any hydrophobic group selected from myristic acid, palmitic acid, stearic acid, polyethylene glycol, and cholesterol; the R2 flanking sequence comprises an amino acid sequence selected from the exemplary sequence set forth in SEQ ID NO: 141 and variants thereof, wherein the C-terminus is amidated; and R3 is selected from any pharmacophore of a small molecule drug, nucleotide, polypeptide, protein, or antibody, or a linker group capable of interacting with any drug of the small molecule drug, nucleotide, polypeptide, protein, or antibody. The vector structure is exemplified by the exemplary structures of SEQ ID NOs: 159-163 and variants thereof.

[0193] In some preferred embodiments herein, the core sequence of the OATP-targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus is selected from the amino acid sequences shown in any one of SEQ ID NOs: 97 and 94, and variants thereof; R1 is a myristic acid group; the R2 flanking sequences are selected from the amino acid sequences shown in SEQ ID NOs: 158 and 140, and variants thereof, and the C-termini are amidated; and R3 can be selected from any pharmacophore of a small molecule drug, nucleotide, polypeptide, protein, or antibody, or a linker group that can interact with any of these drugs. The vector structure is exemplified by SEQ ID NOs: 164-165, and variants thereof.

[0194] In some preferred embodiments herein, the core sequence of the OATP-targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus comprises an amino acid sequence selected from any one of the exemplary sequences shown in SEQ ID NOs: 95-99, 103-110, and variants thereof; R1 is a myristic acid hydrophobic group; R2 is absent; and R3 can be selected from any pharmacophore of a small molecule drug, nucleotide, polypeptide, protein, or antibody, or a linker group capable of interacting with any of the small molecule drugs, nucleotides, polypeptides, proteins, or antibodies. The vector structure is exemplified by the exemplary structures of SEQ ID NOs: 166-178 and variants thereof.

[0195] In one embodiment herein, the OATP targeting vector core sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 97 and variants thereof; R1 is a myristic acid group; the R2 flanking sequence comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 141 and variants thereof, the C-terminus of which is amidated; and R3 is a FITC group.

[0196] The OATP-targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus delivers a pharmacophore or a drug or group linked thereto, and can be used to deliver a pharmacophore or a drug or group linked thereto, including but not limited to mammals, birds, fish, amphibians, and other animals. The mammals include but are not limited to primates, tree shrews, rodents, bats, artiodactyla, carnivora, and other mammals. The birds include but are not limited to ducks, herons, and other avian animals. The primates include but are not limited to hominoid species, New World monkeys, Old World monkeys, and other primates. The hominoid species include but are not limited to humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora.In some preferred embodiments herein, the vector is used for drug delivery to humans, chimpanzees, bonobos, tree shrews, ducks, marmots, Arctic shrews, hamsters, bats, woolly monkeys, capuchin monkeys and squirrel monkeys. In some more preferred embodiments herein, the vector is used for drug delivery to humans.

[0197] The OATP targeted delivery polypeptide vector derived from the surface antigen of the hepadnavirus, wherein the pharmacophore or the drug or group attached thereto, enters systems including but not limited to the digestive system, respiratory system, circulatory system, nervous system, locomotor system, urinary system, reproductive system, endocrine system, and other systems; the pharmacophore or the drug or group attached thereto, wherein the pharmacophore or the drug or group attached thereto, enters organs and tissues including but not limited to the liver, kidney, lung, heart, muscle, fat, brain, nerve, intestine, pancreas, and other organs or tissues. The pharmacophore or the drug or group attached thereto, wherein the pharmacophore or the drug or group attached thereto, preferably enters the digestive system; the pharmacophore or the drug or group attached thereto, preferably enters the liver. In some embodiments herein, the pharmacophore or the drug or group attached thereto, wherein the pharmacophore delivered using the vector, enters systems including but not limited to the digestive system, respiratory system, circulatory system, nervous system, locomotor system, urinary system, reproductive system, endocrine system, and other systems. In some embodiments herein, the pharmacophore of the carrier delivery or the drug or group attached thereto is used to deliver organs and tissues including but not limited to liver, kidney, lung, heart, muscle, fat, brain, nerve, intestine, pancreas and other organs or tissues. In some embodiments herein, the pharmacophore of the carrier delivery or the drug or group attached thereto is used to deliver cells including but not limited to hepatocytes, lung cells, kidney cells, nerve cells, muscle cells, fat cells, blood cells, lymphocytes and other cells. In some preferred embodiments herein, the pharmacophore of the carrier delivery or the drug or group attached thereto is used to deliver into the digestive system and / or liver and / or hepatocytes.

[0198] A hepadnavirus surface antigen-derived peptide that binds to OATP and inhibits its transport function and its application

[0199] Also provided herein is a polypeptide that binds to OATP and inhibits its transport function. The polypeptide is derived from a hepadnavirus surface antigen and retains at least the ability to bind to OATP and inhibit its transport function, i.e., a hepadnavirus surface antigen-derived peptide that binds to OATP and inhibits its transport function, or an "OATP-binding inhibitory peptide derived from a hepadnavirus surface antigen," referred to as an OATP-binding inhibitory polypeptide. Herein, such OATP-binding inhibitory polypeptides can be used to inhibit OATP transport function, regulate the pharmacokinetics of drugs that use OATP as a transporter, and improve the efficacy and safety of these drugs. Therefore, in some embodiments, provided herein is the use of the OATP-binding inhibitory polypeptides described herein in the preparation of a drug for regulating the pharmacokinetics of drugs that use OATP as a transporter, thereby improving the efficacy and safety of these drugs.

[0200] The OATPs that the OATP binding inhibitory polypeptide binds to and inhibits include, but are not limited to, OATP superfamilies from mammals, birds, fishes, amphibians, and other animals. In some embodiments, the OATP superfamily includes, but is not limited to, OATP1, OATP2, OATP3, OATP4, OATP5, OATP6, OATP7, OATP8, OATP9, OATP10, OATP11, and other OATP families. In some embodiments, the OATPs include, but are not limited to, the OATP1B subfamily. In some preferred embodiments, the OATPs include, but are not limited to, OATP1B3, OATP1B1, OATP1B2, and other OATP1B subfamily members.

[0201] The OATP binding inhibitory polypeptide binds to and inhibits OATPs derived from, but not limited to, mammals, birds, fish, amphibians, and other animals. Mammals include, but are not limited to, primates, tree shrews, rodents, bats, artiodactyls, carnivores, and other mammals. Birds include, but are not limited to, ducks, herons, and other avian animals. Primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. The New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. The Old World monkeys include, but are not limited to, crab-eating macaques (Macaca fascicularis) and other Old World monkeys. The rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. The Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include, but are not limited to, rats (Rattus rattus), mice (Mus musculus), and other myomorphs. The Artiodactyla include, but are not limited to, pigs and other even-toed ungulates. The Carnivora include, but are not limited to, dogs and other Carnivora.In some embodiments herein, the polypeptide binds to and inhibits the transport function of OATP1B3, including but not limited to human OATP1B3 (an example sequence can be found in SEQ ID NO: 1, NCBI Reference Sequence: NM_019844), chimpanzee OATP1B3 (an example sequence can be found in NCBI Reference Sequence: XM_016923821), tree shrew OATP1B3 (an example sequence can be found in NCBI Reference Sequence: XM_006155085), cynomolgus monkey OATP1B3 (an example sequence can be found in NCBI Reference Sequence: NM_001283191), Arctic ground hamster OATP1B3 (an example sequence can be found in NCBI Reference Sequence: XM_026391932), rat OATP1B3 (an example sequence can be found in NCBI Reference Sequence: XM_032905480), porcine OATP1B3 (an example sequence can be found in NCBI Reference Sequence: XM_006155085), and porcine OATP1B3 (an example sequence can be found in NCBI Reference Sequence: XM_006155085). Sequence: NM_001315607), OATP1B3 from dog (an example sequence can be found in NCBI Reference Sequence: NM_001166047), and OATP1B3 from other animals mentioned above and their variants. In some preferred embodiments herein, the OATP1B3 bound and modulated in its transport function is OATP1B3 from human primates, rodents, or birds. In some further preferred embodiments herein, the OATP binding inhibitory polypeptide binds to human OATP1B3 and inhibits its transport function. In some embodiments herein, the polypeptide binds to and inhibits the transport function of OATP1B1, including but not limited to human OATP1B1 (an exemplary sequence can be found in SEQ ID NO: 4, NCBI Reference Sequence: NM_006446), bonobo OATP1B1 (an exemplary sequence can be found in NCBI Reference Sequence: XM_003828901), cynomolgus macaque OATP1B1 (an exemplary sequence can be found in NCBI Reference Sequence: NM_001284540), and OATP1B1 and variants thereof from other animals mentioned above. In some preferred embodiments herein, the OATP1B1 bound to and modulated in its transport function is human primate, rodent, or avian OATP1B1. In some further preferred embodiments herein, the OATP binding inhibitory polypeptide binds to human OATP1B1 and inhibits its transport function.In some embodiments herein, the OATP binding inhibitory polypeptide binds to and inhibits the transport function of OATP1B2, including but not limited to rat OATP1B2 (an example sequence can be found in NCBI Reference Sequence: NM_031650), mouse OATP1B2 (an example sequence can be found in NCBI Reference Sequence: NM_020495), and OATP1B2 and variants thereof from other animals mentioned above. In some preferred embodiments herein, the OATP to which the OATP binding inhibitory polypeptide binds and inhibits the transport function is human OATP1B3 and / or human OATP1B1.

[0202] In some embodiments herein, the OATP binding inhibitory polypeptide may bind to and inhibit OATP transport function alone, or may simultaneously bind to and modulate OATP and NTCP transport function. In some embodiments herein, when the polypeptide simultaneously binds to and modulates OATP and NTCP, the NTCP bound and modulated includes, but is not limited to, mammals, birds, fish, amphibians, and other animals. The mammals include, but are not limited to, primates, tree shrews (tupaia), rodents, bats, even-toed ungulates (Artiodactyla), Carnivora, and other mammals. The birds include, but are not limited to, ducks, herons, and other avian animals. The primates include, but are not limited to, hominoid species, New World monkeys, Old World monkeys, and other primates. Hominoid species include, but are not limited to, humans, chimpanzees, gorillas, gibbons, orangutans, and other hominoid species. New World monkeys include, but are not limited to, woolly monkeys, capuchin monkeys, squirrel monkeys, and other New World monkeys. Old World monkeys include, but are not limited to, Macaca fascicularis and other Old World monkeys. Rodents include, but are not limited to, Sciuridae, myomorphs, and other rodents. Sciuridae include, but are not limited to, arctic ground squirrels, ground squirrels, woodchucks, and other Sciuridae. The myomorphs include but are not limited to rats (Rattus rattus), mice (Mus musculus) and other myomorphs. The even-toed ungulates (Artiodactyla) include but are not limited to pigs and other even-toed ungulates.The carnivora animals include, but are not limited to, dogs and other carnivora animals. In some embodiments herein, the NTCPs that are simultaneously bound and regulated include, but are not limited to, NTCP from humans (SEQ ID NO: 7, NCBI Reference Sequence: NM_003049), NTCP from chimpanzees (NCBI Reference Sequence: XM_510035), NTCP from tree shrews (NCBI Reference Sequence: XM_006171503), NTCP from bonobos (NCBI Reference Sequence: XM_003824101), NTCP from capuchin monkeys (NCBI Reference Sequence: XM_032245781), NTCP from squirrel monkeys (NCBI Reference Sequence: XM_003924480), NTCP from cynomolgus monkeys (NCBI Reference Sequence: NM_001283323), NTCP from arctic ground squirrels (NCBI Reference Sequence: XM_026385176), and NTCP from ground hamsters (NCBI Reference Sequence: XM_003824101). Reference Sequence: XM_005322843), NTCP from rat (NCBI Reference Sequence: XM_032908052), NTCP from mouse (NCBI Reference Sequence: NM_001177561), NTCP from porcine (NCBI Reference Sequence: XM_001927695), NTCP from canine (NCBI Reference Sequence: XM_537494), and NTCP and variants thereof from other animals mentioned above. In some preferred embodiments herein, the NTCP protein to which the polypeptide simultaneously binds and regulates is NTCP from human primates, rodents, or birds. In some further preferred embodiments herein, the NTCP protein to which the polypeptide simultaneously binds and regulates is human NTCP.

[0203] In some embodiments herein, the inhibited OATP transport function includes, but is not limited to, OATP transport of the following substrates. Exemplary substrates of OATP1B1 include, but are not limited to, sartans, statins, bile acids and their derivatives, tinibs, platinum drugs, taxanes, chemotherapy drugs, pril drugs, fexofenadine, methotrexate, bosentan, atrasentan, rifampicin, SN-38, irinotecan, cephalosporins, repaglinide, sirolimus, saquinavir, lopinavir, torasemide, and other drugs. Exemplary substrates of OATP1B3 include sartans, statins, bile acids and their derivatives, tinibs, platinum drugs, taxanes, chemotherapy drugs, bosentan, digoxin, hamams, fexofenadine, rifampicin, atrasentan, methotrexate, CCK-8, nafcillin, and other drugs. The statins include, but are not limited to, simvastatin, lovastatin, pravastatin, fluvastatin, atorvastatin, rosuvastatin, cerivastatin, rosuvastatin, pitavastatin, and other statins. The sartans include, but are not limited to, telmisartan, valsartan, irbesartan, olmesartan, valsartan, and other sartans. The platinum drugs include, but are not limited to, cisplatin, carboplatin, oxaliplatin, and other platinum drugs. The paclitaxels include, but are not limited to, paclitaxel, docetaxel, albumin-based paclitaxel, liposomal paclitaxel, and other paclitaxel drugs and their derivatives. Tinib drugs include but are not limited to imatinib, gefitinib, erlotinib, sunitinib, dasatinib, lapatinib, nilotinib, ruxolitinib, crizotinib, icotinib, tofacitinib, bosutinib, cabozantinib, ponatinib, axitinib, ibrutinib, trametinib, ceritinib, apatinib, alectinib, lenvatinib, osimertinib, neratinib, acalabrutinib, brigatinib, baricitinib, and other tinib drugs and their derivatives. Cephalosporins include but are not limited to cefoperazone, cefazolin, cefditoren, cefradine, cefmetazole, cephalexin, and other cephalosporins. The pril drugs include but are not limited to captopril, enalapril, benazepril, lisinopril, ramipril, perindopril, fosinopril, imidapril, cilazapril and other pril drugs and their derivatives.

[0204] The OATP binding inhibitory polypeptide is derived from a surface antigen of a hepadnavirus (Hepadnaviridae) and retains at least the biological activity of binding to OATP and inhibiting OATP substrate transport function. The hepadnavirus includes but is not limited to orthohepadnavirus, avihepadnavirus, metahepadnavirus, herpetohepadnavirus, and other hepadnaviruses. The protohepadnaviruses include but are not limited to protohepadnaviruses that infect humans (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkeys, groundhogs (Woodchuck hepatitis virus, WHV), Arctic ground shrews, ground shrews (Ground squirrel hepatitis virus, GSHV), tree shrews, chimpanzees, bats (Bat hepatitis virus, BHV) and other animals. Example genome sequences can be found in GenBank accession numbers including but not limited to AF046996 (WMHBV), NC_043528 (CMHBV), NC_004107 (WHV), NC_001484 (GSHV), and JX941468 (BHV). The Avihepadnavirus includes but is not limited to Avihepadnavirus that infects ducks (DHBV), herons (HHBV) and other birds, and exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers NC_001344 (DHBV) and NC_001486 (HHBV). The original hepadnavirus (HBV) that infects humans includes but is not limited to genotypes A, B, C, D, E, F, G, H and I genotype HBV, and exemplary genome sequences can be found in, but are not limited to, GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G) and AB179747 (genotype H). In some embodiments herein, the amino acid sequence of the OATP binding inhibitory polypeptide is derived from the amino acid sequence of the viral surface antigen Pre-S region, including but not limited to the exemplary surface antigen Pre-S amino acid sequence SEQ ID NO: 8-22 and variants thereof.In further preferred embodiments herein, the amino acid sequence of the OATP binding inhibitory polypeptide is derived from the amino acid sequence of the human HBV surface antigen Pre-S region, including but not limited to the amino acid sequence of the genotype A, B, C, D, E, F, G, H, and I genotype HBVPre-S region, including but not limited to the exemplary HBV surface antigen Pre-S amino acid sequence SEQ ID NO: 15-22 and variants thereof. In some embodiments herein, the amino acid sequence of the OATP binding inhibitory polypeptide is derived from the amino acid sequence of the human HBV surface antigen Pre-S region, including but not limited to the amino acid sequence of the genotype C genotype HBVPre-S region, including but not limited to the exemplary HBV surface antigen Pre-S amino acid sequence SEQ ID NO: 17 and variants thereof.

[0205] The term "hepadnavirus surface antigen-derived peptide that binds to OATP and inhibits its transport function" or "OATP-binding inhibitory peptide derived from hepadnavirus surface antigen", which may also be referred to as "OATP-binding inhibitory peptide", means that the origin or source of the polypeptide is hepadnavirus, including but not limited to natural, recombinant, synthetic or purified polypeptides, including but not limited to full-length natural HBV surface antigen polypeptides or fragments thereof, and variants of the full-length natural polypeptides or fragments thereof. In some embodiments, the OATP binding inhibitory polypeptide can be derived from any hepadnavirus, including but not limited to orthohepadnavirus (OHV) that infects mammals, avihepadnavirus (AHV) that infects birds, Metahepadnavirus that infects fish, and Herpetohepadnavirus that infects amphibians; in some embodiments, the OATP binding inhibitory peptide can be derived from any hepadnavirus surface antigen; in some embodiments, the OATP binding inhibitory peptide can be derived from any orthohepadnavirus surface antigen; in some embodiments, the OATP binding inhibitory peptide can be derived from any human hepatitis B virus (HBV) surface antigen, including but not limited to HBV surface antigens derived from genotypes A, B, C, D, E, F, G, H, and I; in some embodiments, the OATP binding inhibitory peptide can be derived from the Pre-S1 region of any human hepatitis B virus (HBV) surface antigen, i.e., "OATP binding inhibitory peptide derived from HBVPre-S1" or "HBVPre-S1-derived peptide that binds to OATP and inhibits its transport function."

[0206] The term "OATP-binding inhibitory polypeptide derived from HBVPre-S1" or "HBVPre-S1-derived peptide that binds to OATP and inhibits its transport function" refers to a polypeptide whose origin or source is the HBV surface antigen Pre-S1 region, including but not limited to natural, recombinant, synthetic, or purified polypeptides, including but not limited to the full-length natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, full-length non-natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, non-full-length natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, non-full-length non-natural HBV surface antigen Pre-S1 region, fragments thereof, and variants thereof, and other forms of variants. In some embodiments, the HBVPre-S1-derived peptide that binds to OATP and inhibits its transport function may contain the entire surface antigen pre-S1 region of any HBV genotype surface antigen, starting from amino acid G (glycine) at position 2. In certain embodiments, the HBVPre-S1-derived peptides described herein that bind to OATPs and inhibit their transport function can be derived from the pre-S1 region of the surface antigen of any of HBV genotypes A, B, C, D, E, F, G, H, and I. Exemplary genomic sequences of these HBV genotypes can be found, but are not limited to, GenBank accession numbers KC875260, AY220704, AF461363, AY796030, AB205129, DQ823095, HE981176, and AB179747, and exemplary amino acid sequences of the pre-S regions of their surface antigens can be found, but are not limited to, SEQ ID NOs: 8-22 and variants thereof. In certain embodiments, the HBVPre-S1-derived peptides described herein that bind to OATPs and inhibit their transport function can be derived from the pre-S1 region of the surface antigen of HBV genotype C. The HBVPre-S1 derivative peptide that binds to OATP and inhibits its transport function at least retains the biological activity of binding to OATP and inhibiting its transport function, especially retains the biological activity of binding to OATP1B1, and / or OATP1B2 and / or OATP1B3 and inhibiting their transport function, and especially retains the biological activity of binding to human OATP1B1 and / or human OATP1B3 and inhibiting their transport function.

[0207] In some embodiments, the OATP binding inhibitory peptides described herein may be 10-118 amino acids in length. For example, the length of the polypeptide may be 15-100, 15-80, 20-100, 20-80, 20-60, 25-60, 30-60, 35-60, or 40-60 (including all integers between these ranges) amino acids. In some embodiments, the length of the OATP binding inhibitory peptides described herein may be at least 20 amino acids, for example, at least 25, 30, 35, or 40 amino acids. In some embodiments, the length of the OATP binding inhibitory peptides described herein may be 20, 25, 30, 35, 40, 47, 55, or 60 amino acids. In some embodiments, the length of the OATP binding inhibitory polypeptides described herein may be 47 amino acids. The variants of the OATP binding inhibitory peptides described herein, which have different lengths, retain one or more biological activities associated with the corresponding polypeptides, including at least binding to OATPs (especially OATP1B1 and / or OATP1B3) and inhibiting the transport function of OATPs (especially OATP1B1 and / or OATP1B3).

[0208] In certain embodiments, the OATP-binding inhibitory peptides described herein may be derived from the pre-S1 region of the genotype C HBV surface antigen, and the OATP-binding inhibitory peptides include but are not limited to amino acids 2-119, amino acids 2-69, amino acids 2-59, amino acids 13-119, amino acids 13-88, amino acids 13-72, amino acids 13-67, amino acids 13-59, amino acids 13-52, amino acids 13-47, amino acids 13-42, amino acids 13-37, and amino acids 13-32, and exemplary sequences thereof include but are not limited to SEQ ID NOs: 23-35.

[0209] In some embodiments, the OATP binding inhibitory peptide comprises the amino acid sequence of at least amino acid residues 13-44 at the N-terminus of the pre-S1 region of the genotype C HBV surface antigen as described herein, and variants thereof. In preferred embodiments, the OATP binding inhibitory peptide may be 32-47 amino acid residues in length. In particularly preferred embodiments, the OATP binding inhibitory peptide is the amino acid sequence shown in SEQ ID NO: 30, and variants thereof, comprising at least amino acid residues 1-32 of the exemplary sequence SEQ ID NO: 30, and variants thereof. Preferably, the OATP binding inhibitory peptide comprises at least amino acid residues 1 to 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, or 46 of the exemplary sequence SEQ ID NO: 30, and variants thereof. In some embodiments, the OATP binding inhibitory peptide refers to a fragment containing at least amino acid residues 13-44 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I, and variants thereof, a fragment containing at least amino acid residues 2-33 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and variants thereof, and a fragment containing at least amino acid residues 12-43 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E and G, and variants thereof. In some embodiments, the OATP binding inhibitory peptide comprises a fragment containing at least amino acid residues 13-69 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes A, B, F, H, and I, and variants thereof, a fragment containing at least amino acid residues 2-48 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotype D, and variants thereof, and a fragment containing at least amino acid residues 12-68 at the N-terminus of the pre-S1 region of the HBV surface antigen of genotypes E and G, and variants thereof, exemplary sequences thereof include, but are not limited to, exemplary sequences of SEQ ID NOs: 36-42 and variants thereof.

[0210] As used herein, a "variant" related to a hepadnavirus surface antigen-derived peptide that binds to OATP and inhibits its transport function refers to a variant that differs from a given polypeptide in amino acid sequence but retains the biological activity of binding to OATP and inhibiting its transport function, particularly the biological activity of binding to OATP1B3, and / or OATP1B2, and / or OATP1B1 and inhibiting its transport function, and in particular the biological activity of binding to human OATP1B3 and / or OATP1B1 and inhibiting its transport function. For example, in some embodiments, amino acid substitutions or combinations of amino acid substitutions including, but not limited to, N15D, F25L, G35K, N39E, N46K, F45L, N48H, N48Y, N48K, D50A, H51Q, H51N, E54K, E54D, A55S, N56K, N56D, and Q57K can be introduced into a derivative peptide derived from the amino acid sequence at positions 13-59 of the Pre-S1 region of genotype C HBV. Example sequences include, but are not limited to, SEQ ID NOs: 43-60. The term "variant" also includes homologous polypeptide sequences found in different viral species, strains, or subtypes of the hepadnavirus genotype. HBV is divided into four major serotypes (adr, adw, ayr, and ayw) based on antigenic epitopes on its envelope protein, and into nine genotypes (AI) based on the variability of the entire nucleotide sequence in the genome. Therefore, the term "variant" includes any of these homologous polypeptides found in HBV subtypes. "Variants" may also include polypeptides and variants thereof with naturally occurring flanking amino acid sequences from any of these HBV subtypes added to the N- and / or C-termini. For example, in some embodiments, a polypeptide derived from amino acid sequences 13-59 of the pre-S1 region of the surface antigen of genotype C may be fused with a fragment comprising, but not limited to, at least amino acid residues 2-12 of the N-terminus of the pre-S1 region of the surface antigen of genotypes A, B, F, H, or I, or at least amino acid residues 2-11 of the N-terminus of the pre-S1 region of the surface antigen of genotypes E and G to form a fusion polypeptide, exemplary sequences of which include, but are not limited to, SEQ ID NOs: 61-66. For example, in some embodiments, the polypeptides described herein may contain an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-66, and naturally occurring amino acid sequences flanking the N- and / or C-termini derived from the surface antigen of any of HBV genotypes AI. In some embodiments, the natural flanking amino acid sequence may be derived from HBV surface antigen amino acid sequences including but not limited to GenBank accession numbers KC875260 (genotype A), AY220704 (genotype B), AF461363 (genotype C), AY796030 (genotype D), AB205129 (genotype E), DQ823095 (genotype F), HE981176 (genotype G) or AB179747 (genotype H).For example, the polypeptide described herein may contain the amino acid sequence set forth in SEQ ID NO: 30 and may contain, at its N- and / or C-terminus, a naturally occurring flanking amino acid sequence from the pre-S1 region of HBV genotype C. Alternatively, the polypeptide described herein may contain the amino acid sequence set forth in SEQ ID NO: 30 and may contain, at its N- and / or C-terminus, a naturally occurring flanking amino acid sequence from the pre-S1 region of any one of HBV genotypes A, B, C, D, E, F, G, and H. In some embodiments, the N- and / or C-terminus of the OATP-binding inhibitory peptide described herein may independently contain a naturally occurring flanking amino acid sequence of 1-10 amino acids, such as 1-8, 1-5, or 1-3 amino acids in length (including all integers within these ranges). For example, the OATP-binding inhibitory peptide described herein may contain the amino acid sequence set forth in SEQ ID NO: 30 and may contain, at its N-terminus, a naturally occurring flanking amino acid sequence of 10 amino acids from the pre-S1 region of HBV genotype C. In other words, the polypeptide may comprise amino acids 2-59 of the pre-S1 region of HBV genotype C (SEQ ID NO: 25). As another example, the OATP binding inhibitory peptide described herein may comprise the amino acid sequence set forth in SEQ ID NO: 30 and, at its N-terminus, a naturally occurring flanking amino acid sequence of 9 amino acids from the pre-S1 region of HBV genotype E. In other words, the polypeptide may comprise amino acids 13-59 of the pre-S1 region of HBV genotype C and amino acids 2-11 of the pre-S1 region of HBV genotype E (SEQ ID NO: 63). It should be understood that any of the aforementioned OATP binding inhibitory peptides may have naturally occurring flanking amino acid sequences of any length extending from its N- and / or C-terminus, and the resulting polypeptide retains the biological activity of the original polypeptide in binding to and inhibiting the transport function of OATPs, particularly in binding to and inhibiting the transport function of OATP1B3, and / or OATP1B2, and / or OATP1B1, and in particular in binding to and inhibiting the transport function of human OATP1B3 and / or OATP1B1. "Variants" of the hepadnavirus surface antigen-derived peptides that bind to OATPs and inhibit their transport function also include those having one or more amino acid deletions, substitutions, or insertions, while retaining the biological activity of binding to and inhibiting the transport function of OATPs, particularly the biological activity of binding to and inhibiting the transport function of OATP1B3, and / or OATP1B2, and / or OATP1B1, and especially the biological activity of binding to and inhibiting the transport function of human OATP1B3 and / or OATP1B1. The OATP-binding inhibitory peptide preferably retains the glycine corresponding to amino acid position 13 of the pre-S1 region of genotype C HBV (i.e., the N-terminal glycine of SEQ ID NO: 30).In some embodiments, the OATP binding inhibitory peptides described herein may have one or more naturally occurring mutations in the pre-S1 region of HBV. For example, in some embodiments, the OATP binding inhibitory peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the sequence from the pre-S1 region of HBV. In some embodiments, the OATP binding inhibitory peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to any one of the exemplary amino acid sequences selected from SEQ ID NOs: 23-66. In some embodiments, the OATP binding inhibitory peptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions, substitutions, or insertions relative to the exemplary amino acid sequence of SEQ ID NO: 30. In some embodiments, the OATP binding inhibitory peptides described herein may have 1-3 amino acid deletions, substitutions, or insertions relative to the exemplary amino acid sequence of SEQ ID NO: 30. In certain embodiments, the OATP binding inhibitory polypeptides described herein may have 1-30, such as 1-20, 1-10, 1-8, 1-5, or 1-3 (including all integers within these ranges) amino acid deletions or insertions at the C-terminus relative to any of the exemplary amino acid sequences selected from SEQ ID NOs: 23-66. For example, the OATP binding inhibitory peptides may include, but are not limited to, an amino acid sequence selected from any of the exemplary amino acid sequences of SEQ ID NOs: 31-35. In various embodiments, the present invention includes, but is not limited to, the OATP binding inhibitory polypeptides, as well as polypeptides having at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any of the aforementioned polypeptides. For example, the OATP binding inhibitory peptide may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any of the exemplary sequences in SEQ ID NOs: 23-66. In some embodiments, the OATP binding inhibitory polypeptide may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to any of the exemplary sequences in SEQ ID NOs: 23-66.In some embodiments, the OATP binding inhibitory peptide may comprise an amino acid sequence that is at least about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence exemplified in SEQ ID NO: 30. Variants having a certain sequence identity with the OATP binding inhibitory peptides described herein retain one or more biological activities of the corresponding polypeptide, including at least the biological activity of binding to and inhibiting the transport function of OATPs, particularly the biological activity of binding to and inhibiting the transport function of OATP1B3, and / or OATP1B2, and / or OATP1B1, and particularly the biological activity of binding to and inhibiting the transport function of human OATP1B3 and / or OATP1B1.

[0211] The hepadnavirus surface antigen-derived peptides and variants thereof that bind to OATP and inhibit its transport function are modified at the N-terminus with a hydrophobic group, such as, but not limited to, myristic acid (myr), palmitic acid (plam), stearic acid (stearoyl), cholesterol (chol), oleic acid, linoleic acid, polyethylene glycol (PEG), arachidonic acid, or other hydrophobic groups. In some embodiments, the hydrophobic group can be selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In some embodiments, the hydrophobic group can be myristic acid. In certain embodiments, the OATP binding inhibitory peptides described herein may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, wherein the N-terminus of the OATP binding inhibitory peptide may be modified with a hydrophobic group selected from myristic acid, palmitic acid, stearic acid, and cholesterol. In certain embodiments, the OATP binding inhibitory peptides may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, wherein the N-terminus of the OATP binding inhibitory peptide may be myristoylated. In some embodiments, the OATP binding inhibitory polypeptides described herein may comprise the amino acid sequences set forth in SEQ ID NO:30 and SEQ ID NO:092, wherein the N-terminus of the OATP binding inhibitory peptide may be myristoylated. In some embodiments, the C-terminus of the OATP binding inhibitory polypeptide may be modified. The C-terminal modification may be selected from, but not limited to, amidation (amination), prenylation, and other C-terminal modifications, or may be absent. In some embodiments, the C-terminal modification may be amidation (NH2). For example, but not limited to, the OATP binding inhibitory peptides may comprise the amino acid sequences set forth in SEQ ID NO:30 and SEQ ID NO:60, wherein the C-terminus may be amidated. In some embodiments, the N-terminus of the OATP binding inhibitory peptide may be modified with a hydrophobic group, and / or the C-terminus may be modified with other modifications. In some embodiments, the N-terminus of the OATP binding inhibitory peptide may be modified with a hydrophobic group, and / or the C-terminus may be amidated. In certain embodiments, the OATP binding inhibitory peptide may comprise an amino acid sequence selected from any one of SEQ ID NOs:23-66 and variants thereof, wherein the N-terminus may be modified with a hydrophobic group, and / or the C-terminus may be amidated. For example, but not limited to, the OATP-binding inhibitory peptide may comprise the amino acid sequences set forth in SEQ ID NO: 30 and SEQ ID NO: 60, with the N-terminus modified with a myristic acid (myr), palmitic acid (plam), stearic acid (stearoyl), or cholesterol (chol) hydrophobic group, and the C-terminus modified with amidation, such as the polypeptides set forth in SEQ ID NO: 67-71. In some embodiments, the N-terminus of the OATP-binding inhibitory peptide may be modified with a myristic acid (myr) hydrophobic group, and / or the C-terminus may be modified with amidation.In certain embodiments, the OATP-binding inhibitory peptide may comprise an amino acid sequence selected from any one of SEQ ID NOs: 23-66 and variants thereof, wherein the N-terminus thereof may be modified by myristoylation, and / or the C-terminus thereof may be modified by amidation. For example, but not limited to, the OATP-binding inhibitory peptide may comprise a polypeptide as set forth in SEQ ID NOs: 30, 36-42, or 60, wherein the N-terminus thereof is modified by a myristic acid (myr) hydrophobic group and the C-terminus thereof is modified by amidation, such as the polypeptides as set forth in SEQ ID NOs: 67, 68, or 72-78.

[0212] The hepadnavirus surface antigen-derived peptides encompassed herein bind to OATP and inhibit its transport function, and the polypeptides and variants thereof at least retain the biological activity of binding to OATP and inhibiting its transport function. In some embodiments, the OATP binding inhibitory polypeptides are derived from, but are not limited to, orthohepadnavirus, avihepadnavirus, metahepadnavirus, herpetohepadnavirus, and other hepadnaviruses. In some embodiments, the OATP binding inhibitory polypeptide is derived from, but is not limited to, human (HBV), woolly monkey HBV (WMHBV), capuchin monkey HBV (CMHBV), squirrel monkey, woodchuck hepatitis virus (WHV), arctic shrew, ground shrew (GSHV), tree shrew, chimpanzee, bat hepatitis virus (BHV), and other animal proto-hepadnaviruses, as well as Avihepadnavirus that infects duck HBV (DHBV), heron HBV (HHBV), and other birds. In some embodiments, the OATP binding inhibitory peptide is derived from, but is not limited to, the Pre-S region of the HBV surface antigen of genotypes A, B, C, D, E, F, G, H, and I. In some embodiments, the OATP binding inhibitory peptide is derived from, but is not limited to, amino acids 13-59 of genotype C HBV Pre-S. The OATP binding inhibitory polypeptide is modified with a hydrophobic group at its N-terminus, and may be modified or deleted at its C-terminus to stabilize. In some embodiments, the OATP binding inhibitory peptide is modified with a myristic acid (myr) hydrophobic group, a palmitic acid (plam) group, a stearic acid (stearoyl) group, or a cholesterol (chol) hydrophobic group at its N-terminus, and may be amidated at its C-terminus. In some embodiments, the OATP binding inhibitory peptide is modified with a myristic acid (myr) hydrophobic group at its N-terminus, and may be amidated at its C-terminus. The OATP binding inhibitory polypeptide at least retains the biological activity of binding to OATP and inhibiting its transport function, particularly retains the biological activity of binding to OATP1B1, and / or OATP1B2, and / or OATP1B3, and inhibiting their transport function, and particularly retains the biological activity of binding to human OATP1B1 and / or human OATP1B3, and inhibiting their transport function.In some embodiments, the OATP binding inhibitory peptides specifically include but are not limited to the polypeptides shown in the exemplary sequences SEQ ID NOs: 67-78 and variants thereof; in particular, they include but are not limited to the polypeptides shown in SEQ ID NO: 67 (myrC13-59 NH2) and the exemplary sequence SEQ ID NO: 68 (MyrC13-59NH2 (Q57K)) and variants thereof.

[0213] In some embodiments herein, the OATP binding inhibitory peptide treats or prevents a disease by binding to and inhibiting the transport of an OATP substrate. Specifically, the treatment and prevention of a disease is achieved by administering an optional dose of an OATP binding inhibitory polypeptide. In some embodiments, the method for treating or preventing a disease described herein comprises administering the OATP binding inhibitory peptide and a therapeutic agent, wherein the therapeutic agent is an OATP substrate and is transported via OATP; in some embodiments, the present invention provides the use of the OATP binding inhibitory peptide and the therapeutic agent described herein in the preparation of a medicament for treating or preventing a disease. Preferably, the therapeutic agent is as described in any embodiment herein, particularly a sartan, statin, bile acid derivative, and anti-tumor drug as described below. By binding to OATP and inhibiting the transport of the therapeutic agent by OATP, the pharmacokinetic properties of the therapeutic agent are improved, including but not limited to increasing the plasma concentration of the therapeutic agent, prolonging the duration of the therapeutic agent in the blood, prolonging the half-life of the therapeutic agent, and increasing the concentration of the therapeutic agent in the target tissue or organ, thereby enhancing the therapeutic effect of the therapeutic agent. In some preferred embodiments of the present invention, the therapeutic agent is a sartan drug. The OATP binding inhibitory peptide described herein is used in combination with a sartan drug to inhibit the transport of sartan drugs by OATP and improve the efficacy of sartan drugs.

[0214] In other embodiments, the methods for treating or preventing a disease described herein involve administering an OATP binding inhibitory peptide described herein and a therapeutic agent, wherein the therapeutic agent is an OATP substrate and is cytotoxic. The OATP binding inhibitory peptide described herein binds to OATP and inhibits OATP transport of the therapeutic agent, thereby reducing the drug concentration in cells (particularly hepatocytes), reducing the toxic effects of the therapeutic agent on cells, and improving the safety of the therapeutic agent. These therapeutic agents include, but are not limited to, statins, bile acid derivatives (including, but not limited to, obeticholic acid, ursodeoxycholic acid, and other bile acid derivatives), and other drugs. Statins include, but are not limited to, simvastatin, atorvastatin, rosuvastatin, pravastatin, pitavastatin, lovastatin, atorvastatin, fluvastatin, nuvastatin, cerivastatin, mevastatin, and other statins. In some preferred embodiments herein, the therapeutic agent is a statin. The OATP-binding inhibitory peptide described herein is used in combination with a statin to inhibit the transport of statins by OATP, reduce the hepatotoxicity of statins, and reduce the toxic side effects of statins.

[0215] In other embodiments, the disease treatment or prevention methods described herein are to administer the OATP binding inhibitory peptides described herein and a therapeutic agent, wherein the therapeutic agent is an OATP substrate. While reducing the intracellular concentration of the therapeutic agent and reducing the cytotoxicity of the therapeutic agent, the pharmacokinetic properties of the therapeutic agent are improved, and the therapeutic efficacy is enhanced, thereby achieving both improved efficacy and reduced toxic side effects. In some preferred embodiments herein, the therapeutic agent is an anti-tumor drug. The OATP binding inhibitory peptides described herein are used in combination with the anti-tumor drug to inhibit the transport of OATP to the anti-tumor drug, reduce the hepatotoxicity of the anti-tumor drug, and reduce the toxic side effects of the anti-tumor drug. At the same time, the pharmacokinetic properties of the anti-tumor drug are improved, and the efficacy of the anti-tumor drug is enhanced. The anti-tumor drugs include but are not limited to platinums, paclitaxels, tinibs, methotrexate, and other anti-tumor drugs.

[0216] In other embodiments, the method for treating or preventing a disease described herein is to administer the OATP binding inhibitory peptide described herein. The OATP binding inhibitory polypeptide described herein binds to OATP and inhibits the transport of physiological substrates by OATP in the body, regulating the distribution of physiological substrates, thereby preventing or treating the corresponding disease. The OATP physiological substrates include but are not limited to bile acids, hormones, organic anions, and other OATP physiological substrates. The bile acids include but are not limited to cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), ursodeoxycholic acid (UDCA), lithocholic acid (LCA), and products of these bile acids combined with glycine, taurine, sulfuric acid, glucuronic acid, and other bile acids. In some embodiments herein, the diseases that the OATP binding inhibitory peptide described herein can be used to prevent or treat include but are not limited to liver diseases, metabolic diseases, nervous system diseases, digestive system diseases, fundus diseases, circulatory system diseases, immune system diseases, and other systemic diseases. The liver diseases include but are not limited to fatty liver, alcoholic fatty liver disease, alcoholic steatohepatitis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cholestasis, primary biliary cholangitis, biliary cholangitis, primary biliary cirrhosis (PBC) and other liver diseases related to bile acid, cholesterol, fat, and sugar metabolism and other liver diseases. Metabolic diseases include but are not limited to type 1 diabetes, type 2 diabetes, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperglycemia, high LDL-C, obesity, metabolic syndrome and other metabolic diseases related to cholesterol, fat, and sugar metabolism, and other metabolic diseases. Circulatory system diseases include cardiovascular disease, atherosclerosis, coronary heart disease, hypertension, hyperlipidemia, hypercholesterolemia and other circulatory system diseases.

[0217] In some embodiments, the diseases described herein are various diseases other than liver sites, such as tumors, including but not limited to melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer, Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, chronic lymphocytic leukemia, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, small cell carcinoma, The therapeutic agents used are lung cancer, gastric cancer, colon cancer, malignant pancreatic insulinoma, malignant carcinoid cancer, malignant melanoma, choriocarcinoma, mycosis fungoides, head or neck cancer, osteogenic sarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary system tumors, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, essential thrombocythemia, adrenocortical carcinoma, skin cancer and prostate cancer. Therefore, the therapeutic agent used is a therapeutic drug for these tumors and is transported by OATP.

[0218] A "therapeutically effective amount," "prophylactically effective amount," or "effective amount" of the OATP binding inhibitory peptides and compositions described herein refers to an amount of the OATP binding inhibitory peptide, composition, or therapeutic agent that is effective for treatment or prevention in the prevention or treatment of a disease. The "therapeutically effective amount," "prophylactically effective amount," or "effective amount" may vary depending on the peptide, therapeutic agent, route of administration, disease and its severity, health, age, weight, family history, genetic makeup, stage of pathology, types of prior and concurrent treatments, and other individual characteristics of the subject to be treated. "Treatment" as used herein includes treatment of a subject (e.g., a mammal, such as a human) or cell to alter the current course of the subject or cell. Treatment includes, for...

Claims

1. Use of a substance in the preparation of a medicament for preventing and / or treating infection by hepatitis B virus (HBV) and / or hepatitis D virus (HDV) or diseases related to said infection in animals, wherein: The substance prevents or reduces the interaction between HBV and / or HDV and organic anion transporting polypeptide (OATP) in the animal, and / or prevents or reduces the expression and / or function of OATP in the animal.

2. The use according to claim 1, characterized in that The substance prevents or reduces the interaction between HBV and / or HDV and OATP1B3 and / or OATP1B1 in an animal, and / or prevents or reduces the expression and / or function of OATP1B3 and / or OATP1B1 in an animal.

3. The use according to claim 2, characterized in that The animal is a human, chimpanzee, bonobo, tree shrew, marmot, Arctic ground shrew, ground squirrel, bat, woolly monkey, capuchin monkey or squirrel monkey.

4. The use according to claims 2 and 3, characterized in that The substance prevents or reduces the interaction between HBV and / or HDV and human OATP1B3 and / or human OATP1B1 in the human body, and / or prevents or reduces the expression and / or function of human OATP1B3 and / or human OATP1B1 in the human body.

5. The use according to any one of claims 1 to 4, characterized in that The substance is a polypeptide, antibody, protein, small molecule or polynucleotide that at least retains the activity of acting on OATP, or the activity of acting on the interaction region between HBV or HDV and OATP, or prevents or reduces the expression and / or functional activity of OATP.

6. The use according to any one of claims 1 to 5, characterized in that The substance is used alone, or in combination with an agent that can prevent or reduce the interaction between HBV and / or HDV and NTCP and / or an agent or method that can prevent or reduce the expression and / or function of NTCP, or in combination with other drugs for preventing or treating HBV and / or HDV infection and related diseases.

7. The use according to any one of claims 1 to 6, characterized in that The substance is a polypeptide derived from the surface antigen of hepadnaviridae, and the polypeptide at least retains the biological activity of binding to OATP.

8. The use according to claim 7, characterized in that The polypeptide is derived from the Pre-S1 region of the human HBV surface antigen and has biological activity of binding to human OATP1B3 and / or human OATP1B1. The N-terminus of the polypeptide is modified with a hydrophobic group, and the C-terminus is modified or unmodified.

9. The use according to claim 8, characterized in that The polypeptide is derived from the Pre-S1 region of the surface antigen of human HBV of genotype A, B, C, D, E, F, G, H or I.

10. The use according to claim 8, characterized in that The N-terminus of the polypeptide is modified with fatty acid.

11. The use according to claim 10, characterized in that The N-terminus of the polypeptide is modified with myristic acid.

12. The use according to any one of claims 7 to 11, characterized in that The polypeptide is selected from: a polypeptide derived from amino acids 2-119, 2-69, 2-59, 13-119, 13-88, 13-72, 13-67, 13-59, 13-52, 13-47, 13-42, 13-37 or 13-32 of the Pre-S1 region of the human HBV surface antigen of genotype C, or a variant thereof.

13. The use according to any one of claims 7 to 11, characterized in that The polypeptide is selected from: a polypeptide derived from amino acids 13-59, 13-59, 2-48, 12-58, 13-59, 12-58, 13-59, or 13-59 of the Pre-S1 region of human HBV surface antigen of genotype A, B, D, E, F, G, H, or I, or a variant thereof.

14. The use according to any one of claims 12 and 13, characterized in that The N-terminus of the polypeptide is modified with a fatty acid, and the C-terminus thereof is modified or not.

15. The use according to claim 14, characterized in that The N-terminus of the polypeptide is modified with a fatty acid, and the C-terminus is modified with an amidation. The sequences are shown in SEQ ID NOs: 67-78.

16. The use according to any one of claims 12 to 15, characterized in that The polypeptide is derived from the Pre-S1 region of the C genotype human HBV surface antigen, and its amino acid sequence has one or more substitution mutations selected from the following: N15D, F25L, G35K, N39E, N46K, F45L, N48H, N48Y, N48K, D50A, H51Q, H51N, E54K, E54D, A55S, N56K, N56D and Q57K.

17. A method for establishing a cell or animal model susceptible to HBV and / or HDV, characterized in that: The exogenous SLCO gene is transfected into the cells or animals, and / or the exogenous OATP protein is expressed in the cells or animals, so that the cells or animals that were originally not susceptible to or not easily infected by HBV and / or HDV become susceptible to HBV and / or HDV after being transfected with the SLCO gene and / or expressing the exogenous OATP protein.

18. The method according to claim 17, characterized in that The cells or animals are transfected with exogenous SLCO1B3 and / or SLCO1B1 genes or variants thereof, and / or are caused to express exogenous OATP1B3 and / or OATP1B1 proteins or variants thereof.

19. The method according to claim 18, characterized in that The SLCO1B3 and / or SLCO1B1 exogenous genes are derived from humans, chimpanzees or tree shrews, and the OATP1B3 and / or OATP1B1 exogenous proteins are derived from humans, chimpanzees or tree shrews.

20. The method according to any one of claims 17 to 19, characterized in that SLCO gene was transfected alone or simultaneously with exogenous SLC10A1 gene; exogenous OATP protein was expressed alone or simultaneously with exogenous NTCP protein.

21. The method according to any one of claims 17 to 20, characterized in that The cells are not human embryonic stem cells, but are preferably hepatocytes or liver-derived cell lines transfected with an exogenous SLCO gene and / or expressing an exogenous OATP protein. The animals are non-human mammals, preferably rats, mice, dogs or monkeys transfected with an exogenous SLCO gene or expressing an exogenous OATP protein.

22. The method according to claim 21, characterized in that The cells are hepatocytes or liver-derived cell lines transfected with exogenous SLCO1B3 and / or SLCO1B1 genes or variants thereof derived from humans, chimpanzees or tree shrews, or expressing exogenous OATP1B3 and / or OATP1B1 proteins or variants thereof derived from humans, chimpanzees or tree shrews; the animals are rats, mice, dogs or monkeys transfected with exogenous SLCO1B3 and / or SLCO1B1 genes or variants thereof derived from humans, chimpanzees or tree shrews, or expressing exogenous OATP1B3 and / or OATP1B1 proteins or variants thereof derived from humans, chimpanzees or tree shrews.

23. The method according to claim 22, characterized in that The cells are hepatocytes or liver-derived cells transfected with SLCO1B3 and / or SLCO1B1 exogenous genes or variants thereof from humans, chimpanzees or tree shrews, or expressing OATP1B3 and / or OATP1B1 exogenous proteins or variants thereof from humans, chimpanzees or tree shrews, and simultaneously transfected with SLC10A1 exogenous genes or variants thereof from humans, chimpanzees or tree shrews, or expressing NTCP exogenous proteins or variants thereof from humans, chimpanzees or tree shrews. The animal is a rat, mouse, dog or monkey transfected with an exogenous SLCO1B3 and / or SLCO1B1 gene or a variant thereof derived from a human, chimpanzee or tree shrew, or expressing an exogenous OATP1B3 and / or OATP1B1 protein or a variant thereof derived from a human, chimpanzee or tree shrew, and simultaneously transfected with an exogenous SLC10A1 gene or a variant thereof derived from a human, chimpanzee or tree shrew, or expressing an exogenous NTCP protein or a variant thereof derived from a human, chimpanzee or tree shrew.

24. Cells transfected with an exogenous SLCO gene and / or expressing an exogenous OATP protein, wherein: The cells are not susceptible or not easily infected by HBV and / or HDV before transfection or expression, and become susceptible to HBV and / or HDV after being transfected with the SLCO gene and / or expressing exogenous OATP protein; preferably, the transfection and expression are as described in any one of claims 18-23.

25. A drug or group delivery carrier, characterized in that: The carrier at least retains the biological activity of binding to OATP and delivering the carried drug or group into cells.

26. The delivery vector according to claim 25, characterized in that The OATP is OATP1B3 and / or OATP1B1.

27. The delivery vector according to claim 26, characterized in that The OATP is human OATP1B3 and / or human OATP1B1.

28. The delivery vector of claim 25, wherein The carrier simultaneously binds NTCP.

29. The delivery vector of claim 28, wherein The vector also binds human NTCP.

30. The delivery vehicle of claim 25, wherein The carrier is a polypeptide, a small molecule compound, a protein, an antibody or a nucleic acid, and the carrier binds to the OATP and delivers the carried drug or group into cells and / or organs.

31. The delivery vector of claim 25, wherein The drug or group carried by the carrier is a small molecule drug, nucleic acid, protein, antibody and / or polypeptide, and the drug or group is connected to the backbone of the carrier through covalent bonds and / or non-covalent bonds; wherein the carrier backbone and the drug or group are directly connected or connected through a linker.

32. The delivery vehicle of claim 25, wherein The drug or group carried by the carrier enters the hepatocyte or liver.

33. The delivery vector of claim 32, wherein The drug or group carried by the carrier enters human hepatocytes or human liver.

34. The delivery vector according to any one of claims 31 to 33, characterized in that The carrier is a polypeptide derived from the surface antigen of hepadnavirus, which at least retains the biological activity of binding to OATP and delivering the carried drug or group into cells.

35. The delivery vector of claim 34, wherein The delivery vector has the structure shown in the following formula (I): Wherein G is glycine, the core sequence is an amino acid sequence derived from the surface antigen of the hepadnavirus, X is an amino acid residue having two amino groups or two carboxyl groups or other groups available for linking reactions, R1 is a modification of the N-terminus of the vector, R2 is absent or is a flanking sequence or other group, and R3 is a carried pharmacophore or a linking group that can interact with a drug or group.

36. The delivery vector of claim 35, wherein The core sequence of the delivery vector is derived from the Pre-S region of the hepadnavirus surface antigen.

37. The delivery vector of claim 36, wherein The core sequence of the delivery vector is derived from the Pre-S1 region of the human HBV surface antigen.

38. The delivery vector of claim 37, wherein The core sequence of the delivery vector is derived from amino acids 3-119, 4-119, 3-69, 4-69, 3-59, 4-59, 3-48, 4-48, 14-119, 14-88, 14-72, 14-67, 14-59, 14-56, 14-55, 14-53, 14-48, 14-47, 14-45, 14-42, 14-37 or 14-32 of the Pre-S1 region of the C genotype human HBV surface antigen.

39. The delivery vehicle of claim 37, wherein The core sequence of the delivery vector is derived from amino acids 14-48, 14-48, 3-37, 13-47, 14-48, 13-47, 14-48 or 14-48 of the Pre-S1 region of the human HBV surface antigen of genotype A, B, D, E, F, G, H or I.

40. The delivery vehicle of claim 35, wherein The R1 of the delivery vehicle is a fatty acid group.

41. The delivery vector of claim 40, wherein The R1 of the delivery vehicle is a myristic acid group.

42. The delivery vector of claim 35, wherein The R2 flanking sequence of the delivery vector is derived from amino acids 50-119, 50-88, 50-72, 50-69, 50-67, 58-59, 50-59 or 50-52 of the Pre-S1 region of the C genotype human HBV surface antigen or their variants, wherein the N-terminus of R2 can be modified by amidation.

43. The delivery vehicle of claim 35, wherein The R2 flanking sequence of the delivery vector is derived from amino acids 50-59, 50-59, 39-48, 49-58, 50-59, 49-58, 50-59 or 50-59 of the Pre-S1 region of the human HBV surface antigen of genotype A, B, D, E, F, G, H or I, or a variant thereof, wherein the N-terminus of R2 can be modified by amidation.

44. The delivery vehicle of claim 35, wherein The R3 of the delivery vector is a pharmacophore of a small molecule compound, nucleic acid, polypeptide, protein, antibody or virus, or a linking group that can react with a drug or group such as a small molecule compound, nucleic acid, polypeptide, protein, antibody or virus.

45. The delivery vector of claim 44, wherein R3 of the delivery vector is a small molecule pharmacophore with cell regulatory activity, or a bioactive group derived from an active protein, an active protein-encoding nucleic acid, or a bioactive nucleic acid, wherein the bioactive nucleic acid includes interfering or antisense active protein mRNA, a gene encoding vector, and a viral vector; or, R3 is a linking group that can interact with the small molecule pharmacophore, active protein, or bioactive nucleic acid.

46. ​​The delivery vector according to any one of claims 35 to 45, characterized in that The core sequence of the delivery vector is derived from amino acids 14-48 of the Pre-S1 region of the human HBV surface antigen of genotype C or a variant thereof; R1 is a myristic acid, palmitic acid, stearic acid, cholesterol, or a polyethylene glycol group; the flanking sequence described in R2 is derived from amino acids 50-59 of the Pre-S1 region of the human HBV surface antigen of genotype C, and its C-terminus is amidated; preferably, the amino acid sequence of the delivery vector is as shown in any one of SEQ ID NOs: 228-233.

47. The delivery vector according to any one of claims 35 to 45, characterized in that The core sequence of the delivery vector is derived from amino acids 14-56 of the Pre-S1 region of the C genotype human HBV surface antigen or a variant thereof; R1 is a myristic acid, palmitic acid, stearic acid, cholesterol, or polyethylene glycol group; the flanking sequence described in R2 is derived from amino acids 58-59 of the Pre-S1 region of the C genotype human HBV surface antigen, and its C-terminus is amidated; preferably, the amino acid sequence of the delivery vector is as shown in SEQ ID NO:

234.

48. The delivery vector of claim 46, wherein The core sequence of the delivery vector is derived from amino acids 14-48 of the Pre-S1 region of the C genotype human HBV surface antigen or a variant thereof; R1 is a myristic acid group; the flanking sequence described in R2 is derived from amino acids 50-59 of the Pre-S1 region of the C genotype human HBV surface antigen, and its C-terminus is amidated modified, and R3 is a small molecule pharmacophore; preferably, the amino acid sequence of the delivery vector is as shown in SEQ ID NO:

228.

49. A polypeptide for optimizing the pharmacokinetics of drugs transported via OATP, such as statins, sartans, tinibs, anti-tumor drugs such as platinum drugs or taxanes, or use of a polypeptide in the preparation of a reagent for optimizing the pharmacokinetics of statins, sartans, tinibs, anti-tumor drugs such as platinum drugs or taxanes, wherein: The polypeptide binds to OATP and inhibits its transport function, is derived from the surface antigen of hepadnavirus, and at least retains the biological activity of binding to OATP and inhibiting OATP transport function.

50. The polypeptide or use according to claim 49, characterized in that The polypeptide is derived from the Pre-S1 region of the human HBV surface antigen, has a hydrophobic group modification at its N-terminus, and is modified or unmodified at its C-terminus, and has the activity of binding to and inhibiting the transport function of human OATP1B3 and / or human OATP1B1.

51. The polypeptide or use according to claim 49, characterized in that The polypeptide is derived from the Pre-S1 region of the surface antigen of human HBV of genotype A, B, C, D, E, F, G, H or I.

52. The polypeptide or use according to claim 49, wherein: The N-terminus of the polypeptide is modified with fatty acid.

53. The polypeptide or use according to claim 52, characterized in that The N-terminus of the polypeptide is modified with myristic acid.

54. The polypeptide or use according to any one of claims 49 to 53, characterized in that The polypeptide is selected from: amino acids 2-119, 2-69, 2-59, 13-119, 13-88, 13-72, 13-67, 13-59, 13-52, 13-47, 13-42, 13-37, 13-32 of the Pre-S1 region of the human HBV surface antigen of genotype C or their variants.

55. The polypeptide or use of any one of claims 49-53, derived from amino acids 13-59, 13-59, 2-48, 12-58, 13-59, 12-58, 13-59, or 13-59 of the Pre-S1 region of the human HBV surface antigen of genotype A, B, D, E, F, G, H, or I, or a variant thereof.

56. The polypeptide or use according to any one of claims 54 and 55, which has a fatty acid modification at its N-terminus and has or has no modification at its C-terminus.

57. The polypeptide or use according to claim 56, wherein the N-terminus is modified with a fatty acid and the C-terminus is modified with an amidation, and the sequence is as shown in SEQ ID NOs: 99-110.

58. The polypeptide or use of claim 54, 56 or 57 is derived from the Pre-S1 region of the surface antigen of human HBV of genotype C, and its amino acid sequence introduces N15D, F25L, G35K, N39E, N46K, F45L, N48H, N48Y, N48K, D50A, H51Q, H51N, E54K, E54D, A55S, N56K, N56D, Q57K amino acid mutations or a combination of mutations.

59. The polypeptide or use according to any one of claims 49 to 58, wherein the polypeptide binds to OATP1B3 and / or OATP1B1 and inhibits the transport of statins, sartans, tinibs, anti-tumor drugs such as platinums or paclitaxels.

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