Novel compound and application thereof
阅读说明：本技术 一种新化合物及其应用 (Novel compound and application thereof ) 是由 卢雪琴 穆卓 王圣军 杜艳春 于 2019-12-13 设计创作，主要内容包括：本发明涉及一种新的化合物及其对HBV基因表达抑制的应用。该化合物结构中含有抑制HBV基因表达的干扰核酸、转接点及其末端修饰链。通过末端修饰链,可以在这种siRNA的反义链3’端引入两个或三个的N-乙酰基半乳糖胺,在正义链5’端相应引入两个或一个N-乙酰基半乳糖胺,引入的N-乙酰基半乳糖胺总数量为四。通过体内外药效实验证明,这种新的化合物可以持续高效的抑制HBV基因表达。(The invention relates to a novel compound and application thereof to HBV gene expression inhibition. The structure of the compound contains interfering nucleic acid for inhibiting HBV gene expression, a transfer point and a terminal modification chain thereof. By the end modification of the chain, can be in this siRNA antisense chain 3 'end into two or three N-acetyl galactosamine, in the sense chain 5' end into two or one N-acetyl galactosamine, the total number of the N-acetyl galactosamine introduction is four. In vivo and in vitro efficacy experiments prove that the novel compound can continuously and efficiently inhibit the HBV gene expression.)
1. The compound structurally comprises interfering nucleic acid for inhibiting HBV gene expression, a transfer point and a terminal modification chain thereof, wherein the terminal modification chain consists of a connecting chain D, a connector B, a branched chain L and a liver-targeting specific ligand X, and the terminal modification chain and the interfering nucleic acid are connected through the transfer point R1/R2The structure of the bond is shown as the general formula (I):
wherein:
when n is 1, m is 3; when n is 2, m is also 2;
R1is-NH (CH)2)xCH2-, where x is an integer from 3 to 10;
R2is-NHCH2CH(OH)CH2(OH) -, or a pyrrole or piperidine ring with a primary and secondary alcohol;
the liver targeting specific ligand X is selected from galactose, galactosamine and N-acetylgalactosamine;
the branched chain L is a C3-C18 straight chain containing carbonyl, amido, phosphoryl, oxygen atoms or the combination of the groups;
the linker B is selected from the following structural formulas:
wherein A is1Is C, O, S or NH; r1 is a positive integer from 1 to 15, r2 is an integer from 0 to 5; a. the2Is C, O, S, amino, carbonyl, amido, phosphoryl or thiophosphoryl; the connecting chain D contains C5-C20, and contains amino, carbonyl, amido, oxygen atom, sulfur atom, thiophosphoryl, phosphoryl, cyclic structure or combination of the groups.
2. The compound of claim 1, wherein said interfering nucleic acid includes, but is not limited to, siRNA, miRNA, or Agomir.
3. The compound of claim 2, wherein the interfering nucleic acid is an siRNA.
4. The compound of claim 3, wherein the interfering nucleic acid is siRNA against hepatitis B virus.
5. The compound according to claim 4, wherein the combination of the modified sense strand at the 5 'end of the sense strand and the modified antisense strand at the 3' end of the antisense strand is selected from GBL-01 to GBL-16.
6. The compound according to claim 5, wherein the sense strand of the siRNA obtained by screening has an initiation site of 208, 210, 1522, 1525, 1575, 1576, 1578 or 1580 relative to HBVDNA.
7. The compound of claim 6, wherein said siRNA sequence is selected from the group consisting of:
1) a combined sequence of sense strand SEQ ID NO.1 and antisense strand SEQ ID NO. 2; or
2) A combined sequence of sense strand SEQ ID No.5 and antisense strand SEQ ID No. 6; or
3) A combined sequence of sense strand SEQ ID NO.7 and antisense strand SEQ ID NO. 8; or
4) A combined sequence of sense strand SEQ ID No.9 and antisense strand SEQ ID No. 10; or
5) A combined sequence of sense strand SEQ ID No.13 and antisense strand SEQ ID No. 14; or
6) A combined sequence of sense strand SEQ ID No.17 and antisense strand SEQ ID No. 18; or
7) A combined sequence of sense strand SEQ ID No.27 and antisense strand SEQ ID No. 28; or
8) A combined sequence of sense strand SEQ ID NO.31 and antisense strand SEQ ID NO. 32.
8. A compound according to any one of claims 1-7, selected from GBL-0401 to GBL-0418.
9. Use of a compound according to any one of claims 1-3 for the manufacture of a medicament for the treatment of liver-related diseases including, but not limited to, acute and chronic hepatitis, liver cancer, hereditary liver-derived diseases, cirrhosis, fatty liver, diabetes.
10. Use of a compound according to any one of claims 1 to 8 in the manufacture of a medicament for the treatment of a disease associated with HBV infection, wherein said HBV infection comprises chronic hepatitis b virus infection, acute hepatitis b virus infection.
11. Use according to claim 9 or 10, characterized in that: the liver targeting specific ligand X is specific for asialoglycoprotein receptor (ASGPR) in liver, the related diseases of HBV infection comprise chronic hepatitis B, and the compound can continuously inhibit the expression of HBsAg, HBeAg and HBV DNA of HBV.
12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 8 and a pharmaceutically acceptable excipient, preferably in the form of a subcutaneous injection.
Technical Field
The invention relates to a novel compound and application thereof to HBV gene expression inhibition. The structure of the compound contains interfering nucleic acid for inhibiting HBV gene expression, a transfer point and a terminal modification chain thereof. By the end modification of the chain, can be in this siRNA antisense chain 3 'end into three N-acetyl galactosamine, in the sense chain 5' end into a corresponding two to one N-acetyl galactosamine, the total number of the introduction of N-acetyl galactosamine is four. The drug effect experiments of HepG2 cells and transgenic mice prove that the novel compound can continuously inhibit the expression of HBsAg, HBeAg and HBV DNA of HBV.
Background
RNAi
RNAi (RNA interference) was discovered in 1998 by Andrew Fale (Andrew Z.fire) et al in C.elegans in antisense RNA inhibition experiments, and this process was called RNAi. This finding was evaluated as one of the ten scientific advances in 2001 by the "Science" journal and listed as the first of the ten scientific advances in 2002. Since then, sirnas with RNAi as the mechanism of action have gained wide attention as potential gene therapy drugs, and andru favus and kraig merlo (Craig c. mello) have gained nobel's physiological or medical prize in 2006 due to their contribution in the study of RNAi mechanisms. RNAi is triggered by double-stranded RNA (dsRNA) in many organisms, including animals, plants and fungi, and in the process of RNAi, an endonuclease called "Dicer" cleaves or "dices" long-stranded dsRNA into small fragments 21-25 nucleotides in length. These small fragments, called small interfering RNAs (siRNAs), in which the antisense strand (Guide strand) is loaded onto the Argonaute protein (AGO 2). The AGO2 loading occurs in the RISC-loading complex, which is a ternary complex consisting of the Argonaute protein, Dicer and dsRNA-binding protein (abbreviated TRBP). During loading, the sense strand (Passengerstrand) is cleaved by AGO2 and expelled. AGO2 then uses the antisense strand to bind to mrnas containing fully complementary sequences and then catalyzes the cleavage of these mrnas, resulting in mRNA cleavage that loses the role of the translation template, thereby preventing the synthesis of the protein of interest. After cleavage, the cleaved mRNA is released and the antisense strand loaded RISC-loading complex is cycled for another round of cleavage.
Statistically, about more than 80% of the proteins related to diseases in human bodies cannot be targeted by the conventional small-molecule drugs and biomacromolecule preparations at present, and belong to non-druggable proteins. Gene therapy aiming at treating diseases through functions of gene expression, silencing and the like is considered to be a third generation therapeutic drug following chemical small molecule drugs and biological large molecule drugs, and the therapy realizes treatment of diseases on the gene level and is not limited by unforgeable protein. As the most mainstream type of RNAi technology in gene therapy, RNAi technology treats diseases from mRNA level, and has higher efficiency than treatment of chemical small molecule drugs and biological large molecule drugs at protein level. By utilizing RNAi technology, a sense strand sequence and an antisense strand sequence of siRNA with high specificity and good inhibition effect can be designed according to a specific gene sequence, the single-strand sequences are synthesized through a solid phase, then the sense strand and the antisense strand are paired into siRNA in a specific annealing buffer solution according to the base pairing principle, and finally the siRNA is delivered to a corresponding target spot in vivo through a carrier system, target mRNA is degraded, the function of the target mRNA as a translation template is damaged, and the synthesis of related protein is prevented.
Delivery system for siRNA
sirnas are unstable in blood and tissues, are easily degraded by nucleases, and to improve the stability of sirnas, modifications to the siRNA backbone can be made, but these chemical modifications provide only limited protection from nuclease degradation and may ultimately affect the activity of sirnas. Therefore, a corresponding delivery system is also needed to ensure the safe and efficient crossing of the cell membrane by siRNA. Since the siRNA molecule has large mass, a large amount of negative charges and high water solubility, the siRNA molecule cannot smoothly pass through a cell membrane to reach the inside of a cell.
The basic structure of the liposome is composed of a hydrophilic core and a phospholipid bilayer, the liposome has the phospholipid bilayer similar to a biological membrane and has high biocompatibility, so the liposome becomes the most popular siRNA carrier with the most extensive application once. The liposome-mediated siRNA delivery mainly encapsulates siRNA into liposome, protects the siRNA from being degraded by nuclease, and improves the efficiency of siRNA passing through cell membrane barriers, thereby promoting the absorption of cells. For example, anionic liposomes, pH-sensitive liposomes, immunoliposomes, fusogenic liposomes (fusogenic liposomes) and cationic lipids, etc., although some progress has been made, the liposomes themselves are liable to induce inflammatory reactions, and various anti-histamines and hormones such as cetirizine and dexamethasone must be used before administration to reduce the possible acute inflammatory reactions, so that they are not suitable for all treatment fields in practical clinical application, especially for diseases with long treatment period such as chronic hepatitis b, and the accumulated toxicity which may be generated by long-term use is a potential safety hazard.
Asialoglycoprotein receptor (ASGPR)
The asialoglycoprotein receptor (ASGPR) in the liver is a receptor specifically expressed by liver cells and is a high-efficiency endocytosis type receptor. Since the exposed minor terminal of each glycoprotein is a galactose residue after enzymatically or acid hydrolysis of sialic acid under physiological conditions in vivo, the sugar to which ASGPR specifically binds is a galactosyl group and is also called a galactose-specific receptor. Monosaccharide and polysaccharide molecules such as galactose, galactosamine, and N-acetylgalactosamine have high affinity for ASGPR. The main physiological function of ASGPR is to mediate the elimination of substances such as asialoglycoprotein, lipoprotein and the like in blood, and the ASGPR is closely related to the occurrence and development of liver diseases such as viral hepatitis, liver cirrhosis, liver cancer and the like. The discovery of this property of ASGPR plays an important role in the diagnosis and treatment of Liver-derived diseases (Ashwell G, Harford J, Carbohydrate specific Receptors of the Liver, Ann RevBiochem 198251: 531-554). The liver-derived disease treatment drug containing galactose or galactosamine and derivatives thereof in the structure can be specifically compatible with ASGPR, so that the active liver targeting is realized, and other carrier systems are not needed for delivery.
Disclosure of Invention
The invention relates to a novel compound and application thereof to HBV gene expression inhibition. The structure of the compound contains interfering nucleic acid for inhibiting HBV gene expression, a transfer point and a terminal modification chain thereof. The end modifying strand is connected to the interfering nucleic acid via a transition point. Through the end modification chain, two or three N-acetylgalactosamine can be introduced into the 3 'end of the antisense chain of the siRNA, two or one N-acetylgalactosamine can be correspondingly introduced into the 5' end of the sense chain, and the total number of four N-acetylgalactosamine is a brand new introduction mode.
In one aspect of the present invention, there is provided a compound having a structure comprising an interfering nucleic acid inhibiting HBV gene expression, a transition point and a modified end chain thereof, wherein the modified end chain consists of a connecting chain D, a linker B, a branched chain L and a liver-targeting specific ligand X, and the modified end chain is connected to the interfering nucleic acid via the transition point R1/R2The structure of the bond is shown as the general formula (I):
wherein:
when n is 1, m is 3; when n is 2, m is also 2;
R1is-NH (CH)2)xCH2-, where x may be an integer from 3 to 10;
R2is-NHCH2CH(OH)CH2(OH) -, or other nitrogen-containing structures with both primary and secondary alcohols or primary alcohol alone, which may be a straight chain or branched chain, or a cyclic structure. In some preferred embodiments, R2Can be a pyrrole ring or a piperidine ring with primary and secondary alcohols, and is specifically selected from the following structures:
the liver targeting specific ligand X is selected from galactose, galactosamine and N-acetylgalactosamine;
the branched chain L is a C3-C18 straight chain containing carbonyl, amido, phosphoryl, oxygen atoms or the combination of the groups;
the linker B is selected from the following structural formulas:
wherein A is1Is C, O, S or NH; r1 is a positive integer from 1 to 15, r2 is an integer from 0 to 5; a. the2Is C, O, S, amino, carbonyl, amido, phosphoryl or thiophosphoryl;
the connecting chain D contains C5-C20, and contains amino, carbonyl, amido, oxygen atom, sulfur atom, thiophosphoryl, phosphoryl, cyclic structure or combination of the groups.
In the above technical solution, the interfering nucleic acid includes but is not limited to siRNA, miRNA or ago, preferably siRNA, and more preferably anti-hepatitis b virus siRNA.
The sequence design of the 19mer of the siRNA for HBV RNAi is designed according to HBV cDNA target sequence (GenBank accession # AF 100309.1). These target sequences include the 19mer de1 core regions and the corresponding DNA sequences that are predominantly extended or shifted from these core regions. The aim is to find the optimal effective sequence through the basic target site, and the sequences can be partially or completely matched with the target site and can be applied to treating chronic hepatitis B. The 19mer nucleotide sequence of the target includes two strands, a sense strand (S) and an Antisense Strand (AS). Wherein the 19 th nucleotide (5 '→ 3') of the sense strand and the 1 st nucleotide (5 '→ 3') of the antisense strand can form a base pair according to the warsen-Crick (Watson-Crick) principle. According to this principle, 1 to 19 bases of the sense strand (5 '→ 3') can pair with 19 to 1 bases of the antisense strand (5 '→ 3') and their corresponding bases to form a double strand. The end positions in the duplex may allow one to three unpaired bases to be present. In the invention, HBV siRNA basic sequences can be screened out according to practical application. 3 'or 5' end displacement is carried out according to the basic sequence, and more effective and specific sequences are screened. According to the results of the siRNA structure studies, the optimal choice of single-stranded overhang at the 3' end of the sense or antisense strand is TT, UU, AU or UA, resulting in a variant sequence. Either sense strand can be used to form a duplex with the antisense strand, and both strands must maintain a continuous base pairing of at least 16, 17, 18, 19, 20, 21, 22, or 23. Some of the sequences listed may differ from the target by 1, 2 or 3, and the last base at the 3' end of the sense strand may be U, A or T. The last position at the 3' end of the antisense strand may be U, A or T. According to the principle, the invention screens out the following candidate sequences:
for stability of siRNA in tissues, each monomer of siRNA is modified without affecting or even enhancing activity. The presence of 1, 2 or 3 incompletely paired bases in the sense and antisense strands may be tolerated. The nucleotides may carry various modifying groups, and may be modified in whole or in part. There may be one, more or even all of the thio bearing bases in each strand.
In the compound of the present invention, the modified sense strand and antisense strand are selected from the following sequences:
wherein:
mG, mA, mC and mU are 2 '-methoxy (2' -OMe) modified nucleotides; fG. fA, fC and fU are 2' -fluoro modified nucleotides; s is a phosphorothioate internucleoside linkage and the remaining nucleotidic monomers are linked by phosphodiester linkages. The details are as follows:
g ═ guanylic acid, a ═ adenylic acid, U ═ uridylic acid, C ═ cytidylic acid, dT or T ═ 2' -deoxythymidine;
gs ═ 3 '-thioguanylic acid, As ═ 3' -thioadenosine, Us ═ 3 '-thiouridylic acid, Cs ═ 3' -thiocytidylic acid, dTs or Ts ═ 2 '-deoxy-3' -thiothymidylic acid;
mG ═ 2 '-O-methylguanylic acid, mA ═ 2' -O-methyladenosine, mU ═ 2 '-O-methyluroglycine, mC ═ 2' -O-methylcytidine;
mGs ═ 2 '-O-methyl-3' -thioguanylic acid, mAs ═ 2 '-O-methyl-3' -thioadecanoic acid, mUs ═ 2 '-O-methyl-3' -thiouroglycolic acid, mCs ═ 2 '-O-methyl-3' -thiocytylic acid;
fG 2 '-fluoroguanylic acid, fA 2' -fluoroadenosine, fU 2 '-fluorouridylic acid, and fC 2' -fluorocytidylic acid;
fGs ═ 2 '-fluoro-3' -thioguanylic acid, fAs ═ 2 '-fluoro-3' -thioadenosine acid, fUs ═ 2 '-fluoro-3' -thiouridylic acid, and fCs ═ 2 '-fluoro-3' -thiocytidylic acid.
Further preferably, in some preferred embodiments, in the compound, the modified sense and antisense strands are selected from the following sequences:
the terminal modification chain is composed of a connecting chain D, a connector B, a branched chain L containing a steric stabilization structure and a liver targeting specific ligand X, and the general formula (II) of the terminal modification chain is as follows:
when n ═ 1, general structural formula (II) is:
when n or m is 2, the general structural formula (II) is
When m is 3, the general structural formula (II) is:
the liver targeting specific ligand X may be one or more polysaccharides, polysaccharide derivatives or monosaccharides and monosaccharide derivatives.
Preferably, the liver targeting specific ligand X has the general formula (III):
wherein R is1、R2And R3Hydrogen or a hydroxy protecting group, respectively.
Further preferably, the liver-targeting specific ligand X is selected from galactose, galactosamine, N-acetylgalactosamine or one or more of the following structures:
wherein R is1Selected from OH, NHCOH or NHCOCH3One or two of them.
The branched chain L containing the steric stabilization structure is a linear chain of C3-C18, and the linear chain contains one or more carbonyl groups, amide groups, phosphoryl groups, oxygen atoms or a combination of the groups, and can be selected from one or more of the following structures:
wherein r1 is a positive integer of 1-12, r2 is an integer of 0-20, and Z is H or CH3。
The linker B is selected from the following structural formulas:
wherein A is1Is C, O, S or NH; r1 is a positive integer from 1 to 15, r2 is an integer from 0 to 5; a. the2C, O, S, NH, a carbonyl group, an amide group, a phosphoryl group or a thiophosphoryl group.
Preferably, the linker B is selected from the following structural formulae:
wherein r1, r3, r4 and r5 are positive integers of 1-15 respectively; r6 is a positive integer from 1 to 20, r7 is a positive integer from 2 to 6, and r8 is a positive integer from 1 to 3.
Further preferably, the linker B is selected from the following structures:
the connecting chain D contains C5-C20, and can contain amino, carbonyl, amido, oxygen atom, sulfur atom, thiophosphoryl, phosphoryl, cyclic structure or the combination of the groups.
Preferably, the linking chain D is selected from one of the following structures:
wherein each n is a positive integer from 1 to 20, and each n is the same or different positive integer. p is a positive integer from 1 to 6; s is a positive integer from 2 to 13; r1 and R2 are the same or different substituent groups and have the structural formula of one of the following structures: -H, -CH3、-CH-(CH3)2、-CH2-CH(CH3)2、-CH(CH3)-CH2-CH3、-CH2-C6H5、-C8NH6、-CH2-C6H4-OH、-CH2-COOH、-CH2-CONH2、-(CH2)2-COOH、-(CH2)4-NH2、-(CH2)2-CONH2、-(CH2)-S-CH3、-CH2-OH、-CH(CH3)-OH、-CH2-SH、-CH2-C3H3N2、-(CH2)3NHC(NH)NH2。
Further preferably, the connecting chain D is selected from one of the following structures:
the compound is characterized in that the modified chain at the 5' end of the sense strand is provided with one N-acetylgalactosamine or two N-acetylgalactosamines, and is selected from one of the following structures:
in some preferred embodiments, the modified strand at the 5' end of the sense strand of the compound is selected from the following structural formulae:
the modified strand at the 3' -end of the antisense strand of the compound has two or three N-acetylgalactosamine, and the modified strand has
in some preferred embodiments, the modified strand at the 3' end of the antisense strand of the compound is preferably selected from the following structural formulae:
in some preferred embodiments, the compound wherein the 5 'terminal modified strand of the sense strand in combination with the 3' terminal modified strand of the antisense strand is preferentially one of the structures shown in the following table:
in some preferred embodiments, the compounds of the invention comprise sense strand 5 'ends and antisense strand 3' ends incorporating modified strands as shown in the following table:
in some preferred embodiments, the compounds of the present invention have the following structures as shown in the table:
in another aspect of the present invention, there is provided a use of the compound of the present invention in the preparation of a medicament for treating liver-related diseases, wherein the liver-related diseases include acute and chronic hepatitis, liver cancer, hereditary liver diseases, liver cirrhosis, fatty liver, and diabetes.
In a further aspect, the invention provides the use of a compound of the invention in the preparation of a medicament for the treatment of a disease associated with HBV infection, wherein said HBV infection comprises chronic hepatitis b virus infection, acute hepatitis b virus infection.
Wherein said liver-targeting specific ligand X is specific for asialoglycoprotein receptor (ASGPR) in liver, said HBV infection related disease is chronic hepatitis B, said compound can continuously inhibit the expression of HBsAg, HBeAg and HBV DNA of HBV.
In a further aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable excipient, preferably in the form of a subcutaneous injection.
Compared with the prior art, the invention has the beneficial effects that:
(1) compared to liposome-mediated siRNA delivery: the liposome-mediated siRNA delivery mainly comprises the steps that the liposome encapsulates siRNA into liposome, so that the siRNA is protected from being degraded by nuclease, the efficiency of siRNA passing through cell membrane obstacles is improved, and the absorption of cells is promoted. Although some progress has been made, liposomes such as anionic liposomes, pH-sensitive liposomes, immunoliposomes, fusogenic liposomes (fusogenic liposomes) and cationic lipids, etc., are liable to cause inflammatory reactions, and various anti-histamines and hormones such as cetirizine and dexamethasone, etc. must be used before administration to reduce the possible acute inflammatory reactions, so that they are not suitable for all treatment fields in practical clinical application, especially for diseases with long treatment period such as chronic hepatitis b, and accumulated toxicity generated by long-term use may be a potential safety hazard.
(2) A brand new introduction mode of N-acetylgalactosamine:
compared with siRNA with three N-galactosamine structures in inhibiting HBsAg effect of HBV: the siRNA drugs for treating chronic hepatitis B in the I/II phase at present are ARO-HBV and ALN-HBV 02. ARO-HBV is the introduction of three N-acetylgalactosamine through the linker chain at the 5' end of the sense strand of siARNA; ALN-HBV02 is the 3' end of the sense strand of siRNA, and three N-acetylgalactosamine are introduced by the connecting strand. The galactosamine is introduced into the galactosamine at the positive chain, and three N-acetylgalactosamine are introduced. In the compound provided by the invention, N-acetylgalactosamine with different or same quantity is introduced into the 5' end of the sense strand and the 3' end of the antisense strand of the siRNA at the same time, at present, no report is found that N-acetylgalactosamine is introduced into the 5' end of the sense strand and the 3' end of the antisense strand at the same time, particularly, the introduction of three N-acetylgalactosamine into the 3' end of the antisense strand is a brand new introduction mode, and the embodiment proves that the introduction mode enables the siRNA to have the effect of inhibiting the HBV gene with high efficiency.
Drawings
In order to make the purpose, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 is a high resolution mass spectrum of 5' YICD-01-c 4;
FIG. 2 is a high resolution mass spectrum of 5' YICC-01-c 7;
FIG. 3 is a high resolution mass spectrum of 5' ERCd-01-c 7;
FIG. 4 is a high resolution mass spectrum of 5' ERCc-01-c 4;
FIG. 5 is a high resolution mass spectrum of 3' SANCd-01-c 6;
FIG. 6 is a bar graph of the inhibitory effect on HBsAg in HepG2.215 cells;
FIG. 7 is a bar graph of the inhibitory effect on HBeAg in HepG2.215 cells;
FIG. 8 is a bar graph of the inhibitory effect on HBVDNA in HepG2.215 cells;
FIG. 9 is a bar graph showing the inhibitory effect on HBV gene of transgenic mouse model;
FIG. 10 is a graph showing the inhibitory effect of GBL-0401 on HBV HBsAg in vivo.
Detailed Description
The following examples illustrate some embodiments of the present disclosure, but are not limited thereto. Further, in providing specific embodiments, the inventors contemplate the use of those specific embodiments. For example, compounds having specific chemical structures of the same class or similar, are useful in the treatment of various liver-derived diseases.
Description of the drawings:
the chinese name of DMF is N, N-dimethylformamide;
the Chinese name of HBTU is O-benzotriazole-tetramethyluronium hexafluorophosphate;
DIPEA (DIEA) is named N, N-diisopropylethylamine;
DCM, the chinese name for dichloromethane;
DMAP, the Chinese name being 4-dimethylaminopyridine;
DMT-CL is known by the Chinese name 4,4' -dimethoxytriphenylchloromethane;
THF, chinese name tetrahydrofuran;
the Chinese name of TBTU is O-benzotriazole-N, N, N ', N' -tetramethyluronium tetrafluoroborate;
the Chinese name of DBU is 1, 8-diazabicycloundecen-7-ene;
HOBt, the Chinese name 1-hydroxybenzotriazole;
the chinese name for DCC is dicyclohexylcarbodiimide;
Pd-C is known by its Chinese name as palladium on carbon catalyst;
