CN117105928A - Protease inhibitor and preparation method thereof - Google Patents
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Abstract
The invention provides a protease inhibitor and a preparation method thereof, belonging to the technical field of medicines. The protease inhibitor is a compound structure with the following formula or pharmaceutically acceptable salt thereof. The invention prepares a protease inhibitor compound with strong NS3 protease inhibition effect, improves the pharmacokinetic property, enhances the solubility in an aqueous or lipid environment, improves the oral bioavailability or oral activity, and simultaneously has good NS3 protease inhibition activity, wherein a polyethylene glycol flexible chain enables the compound to keep a certain space configuration and better combine with an active site, thereby playing better inhibition activity.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a protease inhibitor and a preparation method thereof.
Background
Hepatitis C Virus (HCV) is a highly variant positive-strand RNA flavivirus. Patients infected with HCV for more than 6 months will gradually switch to chronic infection with a 50% -85% incidence. And after 20 years of infection, the liver cirrhosis will develop, with the incidence rate of about 10% -15%; can also develop into hepatocellular carcinoma (HCC) with the incidence rate of 1% -7%. The main transmission mode of the hepatitis C virus is blood transmission, and the number of people suffering from hepatitis C after blood transfusion in China is 1/3. In addition, maternal and infant transmission and sexual transmission are also important transmission paths. Clinical manifestations of HCV are insignificant elevations of glutamic pyruvic transaminase, jaundice in few patients, positive HCV antibodies and HCV rna, etc. In recent years, the incidence of hepatitis c has been on the rise worldwide, and the popularity of hepatitis c has been expanding, which has become a serious problem in society and public health.
The pathogenesis of hepatitis C is not yet determined, but in recent years, research shows that the pathogenesis of hepatitis C is similar to that of hepatitis B, and HCV virus can induce human immune response, so that the degeneration and necrosis of liver cells are caused, the liver is directly damaged, and liver lesions are caused. The HCV genome can be translated into multiprotein precursors and then digested to produce structural and nonstructural proteins, including NS2, NS3, NS4A, NS4B, NS5A, NS B, etc. NS3 is a molecule with dual efficacy, having serine-type proteases at the amino-terminus that catalyze cleavage, and nucleoside triphosphates (NTPase) and helicase (helicase) activities at the carboxy-terminus that are essential enzymes for HCV gene translation and replication, and the mechanism of action of protease inhibitors includes blocking NS3 interaction with NS4A, blocking binding of substrates to enzyme active sites, and the like.
Since the first report of this disease in the 80 s of the 20 th century, extensive research has led to the formation of a first generation of therapies for the combination treatment of polyethylene glycol interferon-alpha and ribavirin. This therapy is only effective in about 40% of hepatitis c patients with genotype 1 and exhibits sustained viral inhibition. Recently, direct antiviral Drugs (which act as inhibitors of key enzymes in the viral replication process) have been used as new therapies with better overall response rates (Sheldon, j.; barreiro, p.; soriano, v.expert opin. Invest. Drugs 2007, 16, 1171). One of the targets of these viruses is the NS3 protease. Over the past few years, the pharmaceutical community has struggled to improve HCV therapy, and telaprevir and boceprevir have eventually gained FDA approval. However, some of the properties of these drugs remain to be improved, such as high frequency of administration (both 3 times per day), high mutation rate of HCV leading to faster development of drug resistance, etc.
Polyethylene glycol (polyethylene glycol, PEG) is widely used in the food, pharmaceutical and pharmaceutical industries. Studies of polyethylene glycol solutions have shown that they are soluble in water and organic solvents. The technique of attaching drugs using polyethylene glycol is called "pegylation". Polyethylene glycol technology is now considered as a method of modifying protein drugs, which has the advantage of higher bioavailability, longer half-life, less toxicity and better therapeutic effect. For example, two approved polyethylene glycol interferon products are now approved for the treatment of hepatitis c: polyethylene glycol interferon alpha-2 b interferon (PEG-INTRON) and polyethylene glycol interferon alpha-2 a interferon (PEGSYS). Polyethylene glycols may be linear, branched or designed to vary in molecular weight. PEGylated drugs consisting of polyethylene glycols with molecular weights of 5000-20000Da are generally administered by intravenous route. Small polyethylene glycols (1000 Da or less) can improve pharmacological and pharmaceutical properties for direct use as oral administration.
Disclosure of Invention
The invention aims to provide a protease inhibitor and a preparation method thereof, which have good NS3 protease inhibition activity and better pharmacokinetic properties.
The technical scheme of the invention is realized as follows:
the present invention provides a protease inhibitor which is a compound having the formula I:
a formula I;
wherein n=2-5.
The invention further provides the use of the protease inhibitors described above for the manufacture of a medicament for the treatment or prevention of HCV infection or for the reduction of HCV bioactivity.
As a further improvement of the present invention, the medicament comprises other anti-HCV agents.
The invention further provides the use of the protease inhibitors described above for the preparation of a medicament for inhibiting serine protease activity.
As a further improvement of the present invention, the medicament further comprises a pharmaceutically suitable carrier.
The invention further provides a preparation method of the protease inhibitor, which comprises the following steps:
s1, polyethylene glycol reacts with thionyl chloride to prepare an intermediate A, wherein the structure of the intermediate A is shown as a formula II:
a formula II;
s2, reacting the intermediate A with p-hydroxyacetophenone to prepare an intermediate B, wherein the structure of the intermediate B is shown in a formula III:
formula III;
s3, reacting the intermediate B with bromine water under the catalysis of aluminum chloride to obtain an intermediate C, wherein the structure of the intermediate C is shown as a formula IV:
a formula IV;
s4, reacting ethyl cyanoacetate with hydrogen sulfide to prepare an intermediate D, wherein the structure of the intermediate D is shown as a formula V:
a formula V;
s5, reacting the intermediate D with the intermediate C to prepare an intermediate E, wherein the structure of the intermediate E is shown in a formula VI:
formula VI;
s6, reacting the intermediate E with hydrazine hydrate to prepare an intermediate F, wherein the structure of the intermediate F is shown as a formula VII:
formula VII;
s7, reacting the intermediate F with benzoyl chloride to prepare an intermediate G, wherein the structure of the intermediate G is shown as a formula VIII:
formula VIII;
s8, reacting the intermediate G with phosphorus oxychloride to obtain a product.
As a further improvement of the invention, the molar ratio of the polyethylene glycol to the thionyl chloride in the step S1 is 1:2-2.2, and the structural formula of the polyethylene glycol is as follows:wherein n=2-5, the reaction temperature is room temperature, and the reaction time is 30-50min; in the step S2, alkali is also added, the mol ratio of the intermediate A to the p-hydroxyacetophenone to the alkali is 1:2-2.2:3-5, the reaction temperature is 40-50 ℃, and the reaction time is 2-3h.
As a further improvement of the invention, the molar ratio of the intermediate B to bromine in the step S3 is 1:2-2.2, the reaction temperature is 0-4 ℃ and the reaction time is 1-2h; in the step S4, triethylamine is also added, the mol ratio of the ethyl cyanoacetate to the triethylamine is 1:2-2.2, hydrogen sulfide gas is introduced at the temperature of-20 to-30 ℃, and then the reaction is carried out for 1-2 hours at the temperature of 70-80 ℃; the molar ratio of the intermediate D to the intermediate C in the step S5 is 2-2.2:1, the reaction temperature is 70-80 ℃ and the reaction time is 3-5h.
As a further improvement of the present invention, the molar ratio of intermediate E to hydrazine hydrate in step S6 is 1:20-25, wherein the reaction temperature is 70-90 ℃ and the reaction time is 1-3h; the molar ratio of intermediate F to benzoyl chloride in step S7 is 1:2-2.2, wherein the reaction temperature is 0-4 ℃ and the reaction time is 20-30min; in the step S8, the mol ratio of the intermediate G to the phosphorus oxychloride is 1:5-6, the reaction temperature is 70-90 ℃, and the reaction time is 2-4h.
The invention further protects a drug intermediate B, C, E, F, G, which has a structure shown in the following formulas III, IV, VI, VII and VIII:
formula III;
a formula IV;
formula VI;
formula VII;
formula VIII;
wherein n=2-5.
The preferred dosage of the protease inhibitor is in the range of between 0.1 and about 100mg/kg of body weight per day, more typically between 1 and 50mg/kg of body weight per day, and preferably between 1 and 20mg/kg of body weight per day. The effective dosage range of the pharmaceutically acceptable salts and prodrugs can be calculated based on the weight of the parent compound to be administered. If the salt or prodrug itself exhibits effectiveness, then the effective dose may be estimated as above using the weight of the salt or prodrug, or by other methods known to those skilled in the art.
One mode of administration of the protease inhibitor is oral. The oral compositions will typically include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed in tablets. For the purposes of oral therapeutic administration, the active compounds may be incorporated into excipients and used in the form of tablets, troches or capsules. Pharmaceutically compatible binders and/or adjuvant materials may be included as part of the composition.
If administered intravenously, the preferred carrier is physiological saline or Phosphate Buffered Saline (PBS).
The invention has the following beneficial effects: the invention prepares a protease inhibitor compound with strong NS3 protease inhibition effect, improves the pharmacokinetic property, enhances the solubility in an aqueous or lipid environment, improves the oral bioavailability or oral activity, and simultaneously has good NS3 protease inhibition activity, wherein a polyethylene glycol flexible chain enables the compound to keep a certain space configuration and better combine with an active site, thereby playing better inhibition activity.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that other drawings can be obtained according to these drawings without inventive faculty for a person skilled in the art.
FIG. 1 is a synthetic route diagram of the protease inhibitors of the present invention.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The present invention provides methods for the preparation of certain compounds of the present invention, such as the examples set forth herein. These methods are given for the purpose of illustrating the nature of the preparation process and are not intended to limit the scope of applicable processes. Certain compounds of the present invention may be used as intermediates in the preparation of other compounds of the present invention.
Example 1
Preparation of intermediate a (n=2):
；
0.1mol of ethylene glycol is dissolved in 200mL of dichloromethane, 0.21mol of thionyl chloride is added dropwise at the temperature of 0 ℃, after the addition is finished, the mixture is stirred at room temperature for reaction for 30min, the solvent and the excessive thionyl chloride are removed under reduced pressure, and the product intermediate A is obtained with the yield of 97.2 percent. ESI-MS calculated: c (C) 4 H 9 Cl 2 O (M+H) + 143.01, foundValue: 143.0.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ3.67（t，4H），3.55（t，4H）。
preparation of intermediate B (n=2):
；
0.1mol of intermediate A is dissolved in 200mL of dichloromethane, 0.4mol of triethylamine and 0.21mol of p-hydroxyacetophenone are added, the mixture is heated and refluxed for 3h, the solvent is removed under reduced pressure, and the mixture is separated and purified by column chromatography (petroleum ether: ethyl acetate=1:2) to obtain a product intermediate B with the yield of 92.4 percent. ESI-MS calculated: c (C) 20 H 23 O 5 (m+h) + 343.15, found: 343.1.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ7.75（d，J=6.7Hz，4H），6.88（d，J=6.7Hz，4H），4.12（t，4H），3.78（t，4H），2.55（s，6H）。
preparation of intermediate C (n=2):
；
dissolving 0.1mol of intermediate B in 200mL of anhydrous diethyl ether, cooling to 0 ℃, adding 0.5g of anhydrous aluminum chloride, dropwise adding 0.21mol of bromine water, stirring at 45 ℃ for reaction for 2 hours, removing the solvent under reduced pressure, washing the product, and recrystallizing with ethanol to obtain intermediate C with the yield of 72.2%. ESI-MS calculated: c (C) 20 H 21 Br 2 O 5 (m+h) + 498.97, found: 499.0.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ7.76（d，J=6.5Hz，4H），6.87（d，J=6.5Hz，4H），4.57（s，4H），4.12（t，4H），3.78（t，4H）。
preparation of intermediate D:
dissolving 0.16mol of ethyl cyanoacetate in 100mL of absolute ethyl alcohol, adding 0.32mol of triethylamine, introducing dry hydrogen sulfide gas at the temperature of minus 30 ℃ for 20min at the concentration of 20mL/min, heating to the temperature of 75 ℃ after stopping the gas, stirring for reaction for 2h, cooling to room temperature, and removing the solvent under reduced pressure to obtain a product intermediate D with the yield of 96.2%. ESI-MS calculated: c (C) 5 H 10 NO 2 S (m+h) + 148.04, found: 148.0.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ（q，2H），2.3（s，2H），2.0（br，2H），1.3（t，3H）。
preparation of intermediate E (n=2):
；
dissolving 0.21mol of intermediate D in 400mL of tetrahydrofuran, adding 10g of sodium carbonate, stirring and mixing uniformly, adding 0.1mol of intermediate C, stirring and mixing uniformly, heating and refluxing for 4h, adding saturated sodium bicarbonate solution for quenching reaction, separating liquid, extracting tetrahydrofuran, drying, removing solvent under reduced pressure, and separating and purifying by column chromatography (petroleum ether: ethyl acetate=20:1) to obtain a product intermediate E with the yield of 82.4%. ESI-MS calculated: c (C) 30 H 33 N 2 O 7 S 2 (m+h) + 597.17, found: 597.2.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ7.7（s，2H），7.37（d，J=5.2Hz，4H），6.82（d，J=5.2Hz，4H），4.11-12（m，8H），3.78（t，4H），3.52（s，4H），1.3（s，6H）。
preparation of intermediate F (n=2):
；
0.1mol of intermediate E is dissolved in 150mL of absolute ethanol, 2.5mol of hydrazine hydrate is added, the mixture is heated to 80 ℃, the mixture is stirred for 2h for reaction, the solvent is removed under reduced pressure, and the mixture is washed by ethanol, so that the product intermediate F is obtained, and the yield is 90.1%. ESI-MS calculated: c (C) 26 H 29 N 6 O 5 S 2 (m+h) + 569.16, found: 569.1.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ8.0（br，2H），7.71（s，2H），7.37（d，J=5.4Hz，4H），6.82（d，J=5.4Hz，4H），4.12（t，4H），3.78（t，4H），3.44（s，4H），2.0（br，4H）。
preparation of intermediate G (n=2):
；
0.1mol of intermediate F is dissolved in 100mL of anhydrous tetrahydrofuran, 5G of sodium carbonate and 100mL of water are added, the mixture is stirred and mixed uniformly, 0.21mol of benzoyl chloride is added under ice water bath, the reaction is carried out for 30min, suction filtration and ethanol recrystallization are carried out, and the product intermediate G is obtained with the yield of 93.4%. ESI-MS calculated: c (C) 40 H 37 N 6 O 7 S 2 (m+h) + 777.21, found: 777.2.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ7.92-8.02（m，8H），7.71（s，2H），7.37-7.51（m，10H），6.82（d，J=5.1Hz，4H），4.10（t，4H），3.77（t，4H），3.44（s，4H）。
preparation of product protease inhibitor (n=2):
；
0.1mol of intermediate G is dissolved in 100mL of tetrahydrofuran, 0.55mol of phosphorus oxychloride is added dropwise, the mixture is heated to 80 ℃ and stirred for reaction for 4 hours, the mixture is poured into ice water, stirred and filtered, and the product is obtained by ethanol: recrystallizing the mixed solution with the acetone volume ratio of 2:1 to obtain the product with the yield of 85.7 percent. ESI-MS calculated: c (C) 40 H 33 N 6 O 5 S 2 (m+h) + 741.19, found: 741.2.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ7.7（s，2H），7.48（m，4H），7.32-7.37（m，8H），7.22（m，2H），6.82（d，J=4.9Hz，4H），4.12（t，4H），3.79-3.81（m，8H）。
example 2
Prepared in the same manner as in example 1, except that the starting material was replaced with ethylene glycol (n=3). The structural formula of the product is as follows:
；
ESI-MS calculated: c (C) 42 H 37 N 6 O 6 S 2 (m+h) + 785.21, found: 785.2.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ.7.7（s，2H），7.48（m，4H），7.32-7.37（m，8H），7.22（m，2H），6.82（d，J=4.9Hz，4H），4.11（t，4H），3.77-3.80（m，8H），3.54（s，4H）。
example 3
Prepared in the same manner as in example 1, except that the starting material was replaced with ethylene glycol (n=4). The structural formula of the product is as follows:
；
ESI-MS calculated: c (C) 44 H 41 N 6 O 7 S 2 (m+h) + 829.24, found: 829.2.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ.7.7（s，2H），7.48（m，4H），7.33-7.36（m，8H），7.22（m，2H），6.82（d，J=4.9Hz，4H），4.12（t，4H），3.79-3.82（m，8H），3.53（s，8H）。
example 4
Prepared in the same manner as in example 1, except that the starting material was replaced with ethylene glycol tetraacetal (n=5). The structural formula of the product is as follows:
；
ESI-MS calculated values：C 46 H 45 N 6 O 8 S 2 Actual measurement of (m+h) + 873.27: 873.3.
nuclear magnetic results: 1 H NMR(300MHz，CDCl 3 )δ.7.7（s，2H），7.47（m，4H），7.33-7.35（m，8H），7.21（m，2H），6.83（d，J=4.9Hz，4H），4.11（t，4H），3.78-3.80（m，8H），3.55（s，12H）。
test example 1 measurement of inhibition ratio of NS3/4A protease
Telaprevir, the compound prepared in examples 1-4, was diluted with solvent DMSO to serial dilution concentrations. The NS3/4A protease inhibitor screening model used in the research is a screening model on the established molecular level, and the principle is that the gene 1b type HCV NS3/4A protease is cloned and expressed, a fluorescent agent EDANS and a quencher DABCYL are connected by a section of amino acid sequence which can be recognized and cut by the protease to form a substrate of the protease, and the protease releases the fluorescent agent EDANS by cutting the section of amino acid sequence. Duplicate samples were made for each sample. IC of the active compound is finally calculated according to the koehne modification method 50 。
The results are shown in Table 1.
TABLE 1
As shown in the above table, the protease inhibitors prepared in examples 1 to 4 of the present invention have good inhibitory effect on NS3/4A protease.
Test example 2
The inhibitory activity of viral replication was carried out according to the method in Huh-luc1b replicon cells (Bartensenchlager, et al Science,285, 110, 1999). The compounds prepared in examples 1-4 were prepared as mother liquor with solvent DMSO, diluted to serial dilution concentrations and treated for 72h. Copy number of replicates was measured by bioluminescence and EC was calculated by nonlinear regression 50 Values.
The results are shown in Table 2.
TABLE 2
As can be seen from the above table, the protease inhibitors prepared in examples 1 to 4 of the present invention are high activity NS3 protease inhibitors.
Test example 3
Solubility: measurement of solubility at two pH conditions, 2mg of the protease inhibitors prepared in examples 1 to 4 were added to 1mL of PBS (pH=7.4) or 0.1mol/L hydrochloric acid (pH=2.2) solution. The solution was incubated at room temperature with shaking for 24h. And (3) centrifuging and measuring the concentration in the supernatant. The solubility was calculated by comparison with the amount detected in DMSO at the same concentration. The results are shown in Table 3.
TABLE 3 Table 3
As is clear from the above table, the protease inhibitors prepared in examples 1 to 4 of the present invention have high solubility and high oral bioavailability in rats.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (10)
1. A protease inhibitor, wherein the protease inhibitor is a compound having formula I:
a formula I;
wherein n=2-5.
2. Use of a protease inhibitor according to claim 1 for the manufacture of a medicament for the treatment or prevention of HCV infection or for the reduction of HCV bioactivity.
3. The use according to claim 2, wherein the medicament comprises an additional anti-HCV agent.
4. Use of a protease inhibitor according to claim 1 for the manufacture of a medicament for inhibiting serine protease activity.
5. The use according to any one of claims 2-4, wherein the medicament further comprises a pharmaceutically suitable carrier.
6. A method of preparing the protease inhibitor according to claim 1, comprising the steps of:
s1, polyethylene glycol reacts with thionyl chloride to prepare an intermediate A, wherein the structure of the intermediate A is shown as a formula II:
a formula II;
s2, reacting the intermediate A with p-hydroxyacetophenone to prepare an intermediate B, wherein the structure of the intermediate B is shown in a formula III:
formula III;
s3, reacting the intermediate B with bromine water under the catalysis of aluminum chloride to obtain an intermediate C, wherein the structure of the intermediate C is shown as a formula IV:
a formula IV;
s4, reacting ethyl cyanoacetate with hydrogen sulfide to prepare an intermediate D, wherein the structure of the intermediate D is shown as a formula V:
a formula V;
s5, reacting the intermediate D with the intermediate C to prepare an intermediate E, wherein the structure of the intermediate E is shown in a formula VI:
formula VI;
s6, reacting the intermediate E with hydrazine hydrate to prepare an intermediate F, wherein the structure of the intermediate F is shown as a formula VII:
formula VII;
s7, reacting the intermediate F with benzoyl chloride to prepare an intermediate G, wherein the structure of the intermediate G is shown as a formula VIII:
formula VIII;
s8, reacting the intermediate G with phosphorus oxychloride to obtain a product.
7. The method according to claim 6, wherein the molar ratio of polyethylene glycol to thionyl chloride in step S1 is 1:2-2.2, and the polyethylene glycol has the following structural formula:wherein n=2-5, the reaction temperature is room temperature, and the reaction time is 30-50min; in the step S2, alkali is also added, the mol ratio of the intermediate A to the p-hydroxyacetophenone to the alkali is 1:2-2.2:3-5, the reaction temperature is 40-50 ℃, and the reaction time is 2-3h.
8. The preparation method according to claim 6, wherein the molar ratio of the intermediate B to bromine in the step S3 is 1:2-2.2, the reaction temperature is 0-4 ℃ and the reaction time is 1-2h; in the step S4, triethylamine is also added, the mol ratio of the ethyl cyanoacetate to the triethylamine is 1:2-2.2, hydrogen sulfide gas is introduced at the temperature of-20 to-30 ℃, and then the reaction is carried out for 1-2 hours at the temperature of 70-80 ℃; the molar ratio of the intermediate D to the intermediate C in the step S5 is 2-2.2:1, the reaction temperature is 70-80 ℃ and the reaction time is 3-5h.
9. The method according to claim 6, wherein the molar ratio of the intermediate E to the hydrazine hydrate in the step S6 is 1:20-25, wherein the reaction temperature is 70-90 ℃ and the reaction time is 1-3h; the molar ratio of intermediate F to benzoyl chloride in step S7 is 1:2-2, reacting at 0-4 ℃ for 20-30min; in the step S8, the mol ratio of the intermediate G to the phosphorus oxychloride is 1:5-6, the reaction temperature is 70-90 ℃, and the reaction time is 2-4h.
10. A pharmaceutical intermediate B, C, E, F, G, characterized by having the structure of formula III, formula IV, formula VI, formula VII, formula VIII:
formula III;
a formula IV;
formula VI;
formula VII;
formula VIII;
wherein n=2-5.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003034684A (en) * | 2001-06-29 | 2003-02-07 | Biofor Inc | Antifungal agent of new kind |
| US20030181363A1 (en) * | 2002-01-30 | 2003-09-25 | Boehringer Ingelheim (Canada) Ltd. | Macrocyclic peptides active against the hepatitis C virus |
| WO2004108719A1 (en) * | 2003-06-04 | 2004-12-16 | B & C Biopharm Co., Ltd. | 2-[2-(4-morpholino)ethylamino]pyridine derivatives, method for preparing thereof and antiviral pharmaceutical composition comprising the same |
| ES2240446T3 (en) * | 2000-04-03 | 2005-10-16 | Vertex Pharma | SERINE PROTEASE INHIBITORS, PARTICULARLY THE NS3 PROTEASE OF HEPATITIS VIRUS C. |
| US20070004723A1 (en) * | 2004-02-23 | 2007-01-04 | Dane Goff | Heterocyclic compounds having an oxadiazole moiety and hydro isomers thereof |
| US20080181868A1 (en) * | 2006-06-06 | 2008-07-31 | Ying Sun | Macrocylic oximyl hepatitis c protease inhibitors |
| CN101679325A (en) * | 2007-02-23 | 2010-03-24 | 吉里德科学公司 | The modulators of pharmacokinetic properties of therapeutical agent |
| AU2012200209A1 (en) * | 2004-10-01 | 2012-02-02 | Vertex Pharmaceuticals Incorporated | HCV NS3-NS4A Protease Inhibition |
| CN102413827A (en) * | 2009-04-25 | 2012-04-11 | 弗·哈夫曼-拉罗切有限公司 | Methods for improving pharmacokinetics |
| CN102482269A (en) * | 2009-12-04 | 2012-05-30 | 财团法人卫生研究院 | Proline derivatives |
| US20130028859A1 (en) * | 2011-02-07 | 2013-01-31 | Bristol-Myers Squibb Company | Hepatitis C Virus Inhibitors |
-
2023
- 2023-08-22 CN CN202311056005.1A patent/CN117105928B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2240446T3 (en) * | 2000-04-03 | 2005-10-16 | Vertex Pharma | SERINE PROTEASE INHIBITORS, PARTICULARLY THE NS3 PROTEASE OF HEPATITIS VIRUS C. |
| JP2003034684A (en) * | 2001-06-29 | 2003-02-07 | Biofor Inc | Antifungal agent of new kind |
| US20030181363A1 (en) * | 2002-01-30 | 2003-09-25 | Boehringer Ingelheim (Canada) Ltd. | Macrocyclic peptides active against the hepatitis C virus |
| WO2004108719A1 (en) * | 2003-06-04 | 2004-12-16 | B & C Biopharm Co., Ltd. | 2-[2-(4-morpholino)ethylamino]pyridine derivatives, method for preparing thereof and antiviral pharmaceutical composition comprising the same |
| US20070004723A1 (en) * | 2004-02-23 | 2007-01-04 | Dane Goff | Heterocyclic compounds having an oxadiazole moiety and hydro isomers thereof |
| AU2012200209A1 (en) * | 2004-10-01 | 2012-02-02 | Vertex Pharmaceuticals Incorporated | HCV NS3-NS4A Protease Inhibition |
| US20080181868A1 (en) * | 2006-06-06 | 2008-07-31 | Ying Sun | Macrocylic oximyl hepatitis c protease inhibitors |
| CN101679325A (en) * | 2007-02-23 | 2010-03-24 | 吉里德科学公司 | The modulators of pharmacokinetic properties of therapeutical agent |
| CN102413827A (en) * | 2009-04-25 | 2012-04-11 | 弗·哈夫曼-拉罗切有限公司 | Methods for improving pharmacokinetics |
| CN102482269A (en) * | 2009-12-04 | 2012-05-30 | 财团法人卫生研究院 | Proline derivatives |
| US20130028859A1 (en) * | 2011-02-07 | 2013-01-31 | Bristol-Myers Squibb Company | Hepatitis C Virus Inhibitors |
Non-Patent Citations (2)
| Title |
|---|
| COLUMBUS, OHIO, US: "《STN检索报告 US REGISTRY》", REGISTRY[ONLINE], 22 July 1998 (1998-07-22), pages 1 * |
| 刘俊芝: "新型HCV_NS3_NS4A蛋白酶抑制剂的设计与合成", 中国优秀博硕士学位论文全文数据库（硕士）工程科技Ⅰ辑, no. 03, 15 September 2007 (2007-09-15), pages 016 - 115 * |
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