JP2005511749A - Use of phosphonate nucleotide analogues to treat hepatitis B virus infection - Google Patents

Abstract

  2-Amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- () for oral administration to treat hepatitis B virus infection in infected patients A pharmaceutically acceptable composition comprising 4-methoxyphenylthio) purine (LY582563) is provided. The composition is in unit dosage form containing from about 2.5 to about 20 mg of the purine per unit dose. The composition is adapted for oral administration and is preferably in the form of a tablet or capsule and can be administered in single or multiple doses. However, the total daily dose of the purine is about 2.5 mg to about 20 mg per patient per day. These compositions are particularly advantageous for lowering plasma HBV DNA levels in human patients with chronic HBV infection or ameliorating symptoms, conditions, or disorders caused by hepatitis B virus.

Description

  The present invention relates to 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl]-for treating hepatitis B virus (HBV) infection in human patients. It relates to a new dosage and dosing schedule for the administration of 6- (4-methoxyphenylthio) purine (LY582563). The present invention is particularly advantageous for reducing plasma HBV DNA levels in patients infected with HBV and ameliorating symptoms, conditions, or disorders associated with human HBV infection.

  This application is a U.S. provisional application No. 60 / 338,242 filed on Dec. 7, 2001, No. 60 / 372,013 filed on Apr. 11, 2002, and No. 60 filed on May 16, 2002. No. 381/123, the benefit of priority, all of which are hereby incorporated by reference.

  Hepatitis B is caused by infection with hepatitis B virus, a partial double-stranded DNA virus of the Hepadnaviridae family, and is sometimes a fatal liver disease. Approximately 350 million people worldwide are chronically infected with HBV, and it is estimated that approximately 1 million people die each year as a direct result of HBV-induced cirrhosis or liver cancer.

  HBV is a hepatophilic virus that replicates in the liver and causes acute and chronic liver disease, including liver fibrosis, cirrhosis, inflammatory liver disease and liver cancer, and may be fatal in some of these patients (WKJoklik, Virology, 3rd edition, Appleton & Lange, Norwalk, Conneticut, 1988). The virus is a blood-borne pathogen that is transmitted in a manner similar to human immunodeficiency (HIV) when exposed to infectious fluids. However, HBV is much more susceptible to infection than HIV. Effective vaccines are available, but there is no therapeutic value for those already infected with the virus. It is estimated that 15-20% of chronically infected individuals develop cirrhosis or another chronic disorder from HBV infection. For patients with compensated cirrhosis, survival is 84% after 5 years and 68% after 10 years. In carriers of decompensated cirrhosis, the 5-year survival rate is only 14% (Lok et al. (2001) Hepatology 34: 1225-1241).

  α-interferon therapy is available for the treatment of chronic hepatitis B for the time being. However, less than 40% of patients are effective and have side-effects that limit dosage, such as flu-like symptoms, weight loss, depression, and cytopenias. Lamivudine and adefovir are novel compounds currently under development that are used for the treatment of HBV. Unfortunately, the emergence of lamivudine-resistant virus has occurred in up to 70% of patients after 4 years. Therefore, the development of improved and effective anti-HBV anti-hepatitis therapies is necessary and a top priority (Locarnini et al. (1996) Antiviral Chemistry & Chemotherapy 7 (2): 53-64).

  LY582563 (2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine) is a recently discovered phosphonate nucleotide It is an analog and is currently under development as an anti-HBV drug (see US Pat. No. 5,840,716). LY582563 is a derivative of 9- [2- (phosphonylmethoxy) ethyl] adenine (PMEA; adefovir) with improved oral absorption and antiviral activity. In vitro, LY582563 exhibits higher activity against HBV than lamivudine or PMEA. Although some of the efficacy of LY582563 in the treatment of HBV infection is known, the optimization of treatment with this agent has considerable therapeutic value.

U.S. Provisional Patent Application No. 60 / 338,242 U.S. Provisional Patent Application No. 60 / 372,013 US Provisional Patent Application No. 60 / 381,123 U.S. Pat.No. 5,840,716 W.K.Joklik, Virology, 3rd edition, Appleton & Lange, Norwalk, Conneticut, 1988 Lok et al. (2001) Hepatology 34: 1225-1241. Locarnini et al. (1996) Antiviral Chemistry & Chemotherapy 7 (2): 53-64.

  That is, the present invention is directed to a novel method of treating human HBV infection, including a novel pharmaceutical composition comprising LY582563 and an optimal dosing regimen for administration of LY582563 to treat HBV-infected patients. To do.

  That is, in the first aspect, the present invention provides about 2.5 mg to about 20 mg of 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] per dosage unit. A pharmaceutical composition for oral administration in dosage unit form comprising -6- (4-methoxyphenylthio) purine and a pharmaceutically acceptable carrier, diluent or excipient is provided.

  The composition comprises the purine in an amount of about 2.5 mg, about 5 mg, about 10 mg, about 15 mg, or about 20 mg per dosage unit, or in an amount of about 5 mg to about 10 mg, or about 7.5 mg to about 10 mg per dosage unit. In the form of tablets or capsules. In each case, the upper limit of these ranges may extend to about 12 or about 12.5 mg per dosage unit.

  In another aspect, the invention provides 2-amino-9- [2- [bis (2,2,2-trimethyl) for the preparation of a medicament for treating a human patient suffering from hepatitis B virus infection. Fluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine, wherein the medicament is prepared for oral administration and the medicament is in dosage unit form, Use is provided comprising from about 2.5 mg to about 20 mg of the purine.

  The medicament comprises the purine in an amount of about 2.5 mg, about 5 mg, about 10 mg, about 15 mg, or about 20 mg per dosage unit, or about 5 mg to about 10 mg, or about 7.5 mg to about 10 mg per dosage unit. It can be in the form of a tablet or capsule containing. In each case, the upper limit of these ranges may extend to about 12 or about 12.5 mg per dosage unit.

  In another aspect, the invention provides 2-amino-9- [2- [bis (2,2,2-trimethyl) for treating a human patient suffering from hepatitis B virus infection. There is provided a method comprising administering fluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine in a total amount of about 2.5 mg to about 20 mg of the purine per day.

In this method, the purine can be administered to the patient for a period sufficient to lower the plasma HBV / DNA amount of the patient as compared to the plasma HBV / DNA amount of the patient before the purine is administered. Preferably, the amount of plasma HBV · DNA in the patient is reduced to at least about 10 ⁴ copies / mL compared to the amount of plasma HBV · DNA in the patient prior to administration of the purine. Alternatively, the purine is administered to a patient for a period of time sufficient to treat or ameliorate a symptom, condition, or disorder caused by the patient's hepatitis B virus, such as liver fibrosis, cirrhosis, inflammatory liver disease, or liver cancer can do.

  In any of the foregoing methods, the purine can be administered to the patient in the form of a pharmaceutically acceptable oral composition that may be a tablet or capsule. Further, the period for administering the purine may be days, weeks, months, or years, and the total amount of purine is about 2.5 mg, about 5 mg, about 10 mg, about 15 mg, or about 20 mg per day. Or in an amount of about 5 mg to about 10 mg per day, or about 7.5 mg to about 10 mg per day. In each case, the upper limit of these ranges may extend to about 12 or about 12.5 mg per day. These purine amounts can be administered in a single dose or in divided doses totaling a total amount per day.

  Further scope of applicability of the present invention will become apparent from the detailed description provided hereinafter. However, since the detailed description will reveal various modifications and variations within the spirit and scope of the invention, the detailed description and specific examples illustrate the preferred embodiments of the invention. However, it should be understood that this is merely an example.

  The above and other aspects, features and advantages of the present invention will be better understood from the following detailed description, taken in conjunction with the accompanying drawings, which are illustrative only and not limiting of the present invention.

  The following detailed description of the invention is provided to assist those skilled in the art in practicing the invention. Even so, modifications and variations of the embodiments discussed herein can be made by those skilled in the art without departing from the spirit or scope of the discovery of the present invention, so the following detailed description is provided by the present invention. Should not be considered excessively limiting.

  The contents of all references cited herein are hereby incorporated by reference.

  Current treatment choices for chronic HBV infection are limited in number and clinical utility due to the problems of tolerability and the emergence of resistant virus variants. LY582563, shown below, is a new purine nucleotide analog prodrug that has shown potent activity against HBV in preclinical studies (Ono-Nita et al. (2002) Antimicrob. Agents Chemother. 46 (8): 2602 -5 pages; Wise et al. (2002) J. Gastroenterol. Hepatol. 17 (suppl): A46).

  Regarding oral administration of LY582563, US Pat. No. 5,840,716 teaches that the clinical dosage for adults may be 0.1 mg to 500 mg / kg of compound per day, preferably 1 mg to 50 mg / kg per day. For adults weighing 68 kg (about 150 pounds), these ranges are from about 6.8 mg to about 34 g per day, preferably from about 68 mg to about 3.4 g per day. The '716 patent points out that these dosages may be appropriately increased or decreased depending on age, condition or symptom, or the presence or absence of co-administered drugs. In addition, US Pat. No. 5,840,716 teaches that the above daily dose may be administered once a day or divided into two or more times a day at an appropriate interval. . Finally, continuous administration can be performed at intervals of several days.

  Determining the safe and effective dose of any new drug and the appropriate treatment regimen that uses the drug is an empirical process. This process generally requires treating sick patients one or more times daily with different doses of drugs for different periods of time and monitoring their effectiveness with accepted diagnostic methods And In the case of HBV, these methods include plasma or tissue levels of various antigens, including HBV DNA or parts of the virus, or patient responses such as antibody formation (eg, HBe antibody) or alanine aminotransferase (ALT) levels. Measurements of biochemical improvements, such as declines, can be included. In addition, the effectiveness of treatment can be measured by monitoring the recovery of symptoms, conditions, or disorders caused by the virus. In the case of HBV, these include fatigue, eating disorders, jaundice, liver fibrosis and inflammation, cirrhosis, and liver cancer (hepatocellular carcinoma). However, in addition to demonstrating efficacy, it must also demonstrate acceptable safety, including acceptable toxicity. Toxicity seen with other HBV antiviral drugs includes mitochondrial toxicity, nephrotoxicity, and bone marrow toxicity. The dose and treatment regimen of new drugs, and the relationship between efficacy and safety can only be determined by human clinical trials. Therefore, the determination of dosage and treatment plan parameters cannot be predicted or inferred, and the results are necessarily new and non-obvious. Special attention should be given to the selection of treatment parameters so that they are effective in the treatment of certain diseases while at the same time causing no other harm to the patient. Because of these problems, the findings regarding the use of LY582563 in the treatment of HBV infection in human patients as disclosed herein are surprising and can be predicted prior to conducting this study There should have been no. These results provide a rationale for safe and effective dosing and treatment planning using LY582563 for the treatment of HBV.

  The studies disclosed herein have determined the optimal dosage and dosing regimen for treating patients with HBV infection using LY582563, thereby maximizing the safety and therapeutic benefits obtained with this antiviral agent. Was started to become. Briefly, LY582563 was administered to chronic HBV-infected patients for 28 days in a randomized placebo-controlled dose escalation study. The dose of LY582563 included 2.5 mg, 5 mg, and 10 mg QD or BID, and 20 mg QD. A placebo was also used. HBV DNA was measured in treated patients during dosing and during a follow-up period of up to 12 weeks. Safety data, such as clinical signs (e.g. blood pressure, pulse, respiratory rate, and general physical findings), adverse events (e.g. epigastric pain, diarrhea, nausea, headache, fatigue, and alopecia), ECG, blood tests Blood chemistry measurements (e.g., aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate, blood urea nitrogen (BUN), creatinine, and bilirubin), and sensitive urine markers for renal tubular toxicity (Lactate dehydrogenase (LDH), urine creatinine, and urinary β-N-acetylglucosaminidase (β-NAG)) were also measured. The data obtained demonstrates that LY582563 exhibits a good safety profile and effective HBV antiviral activity in chronically infected patients, for long-term safe and effective use of this new HBV antiviral drug Suggests optimal dosage levels and dosing schedules. According to this result, a suitable dose that balances both safety and efficacy of LY582563 is about 2.5 mg to about 20 mg per patient per day. These dosages may have to be administered to patients over a period of days, weeks, months or years to remedy or control the undesired pathogenic consequences of HBV infection.

Dosage plan for treatment of HBV with LY582563
Oral regimens for treating patients suffering from HBV with LY582563 generally include the patient's age, weight, sex, dietary restrictions and medical condition, infectious virus genotype, severity of infection, and activity of LY582563, Selection is based on a variety of factors, including pharmacological issues such as efficacy, pharmacokinetics and toxicity profiles. Administration of LY582563 generally results in recovery or complete elimination of one or more signs, symptoms, conditions, or disorders present in the patient until the virus titer reaches an acceptable amount or due to hepatitis B virus infection And should continue for hours, days, or weeks to months or years until it is shown that the infection has been controlled or eradicated.

Respect reduction in plasma HBV · DNA amount, long term goal is a matter of current discussion, the part researchers in this field, the desired amount has proposed to be less than ¹⁰⁴ copies / mL. The duration of this HBV / DNA amount has not yet been defined.

  Representative signs of HBV infection other than plasma HBV / DNA levels include, but are not limited to, liver fibrosis, cirrhosis, inflammatory liver disease, and liver cancer. Hollinger et al. (2001) Fields Virology, 4th edition, volume 2, David M. Knipe et al., Ed., "Hepatitis B Virus" chapter 87, pages 2971-3036, Lippincott, Williams, & Wilkins, Philadelphia, PA, USA Is outlined. In most parts of the world, liver fibrosis is diagnosed by liver biopsy, and in many Asian countries liver biopsy is not usually done, and doctors usually make speculative diagnoses using clinical measurements .

Patients who have been treated with LY582563 can measure the amount of HBV / DNA in the serum by, for example, slot-blot, dot-blot, or PCR to determine the effectiveness of the treatment, or serum In addition, it can be monitored according to a conventional method by measuring HBV antigens such as HBV surface antigen (HBsAg) and HBVe antigen (HBeAg) in tissues. Methods for that are described in Hoofnagle et al. (1997) New Engl. Jour. Med. 336 (5): 347-356; FD Hollinger (1996) Fields Virology, 3rd edition, Volume 2, Bernard N. Fields et al., Ed. `` Hepatitis B Virus '', Chapter 86, pages 2738-2807, Lippincott-Raven, Philadelphia, Pennsylvania, USA; and Hollinger et al. (2001) Fields Virology, 4th Edition, Volume 2, David M. Knipe et al., Ed. , "Hepatitis B Virus", chapter 87, pp. 2971-3036, in particular pages 2989-3091, Lippincott, Williams, & Wilkins, Philadelphia, PA, USA and references cited therein. In chronic hepatitis B, remission is the disappearance of HBV viral DNA, ie, for example, an amount of ≧ 10 ⁵ genomes per ml of serum can be detected, or HBeAg can be detected from serum despite the continued presence of HBsAg It is characterized by a decrease to an undetectable amount when measured by a simple hybridization test. Following these serological events, the biochemical and histological characteristics of the disease are improved. The end point of successful treatment in most antiviral therapy trials is the disappearance of HBeAg and viral DNA from serum. In patients who have lost the e antigen, remission usually persists, resulting in an inactive HBsAg carrier status. Many Caucasian patients who are mostly ill during adulthood and are receiving interferon therapy eventually become HBsAg negative (Hoofnagle et al. (1997) New Engl. Jour. Med. 336 (5): 347-356). Is outlined). However, this does not represent about 90% of the global HBV population. Ongoing analysis of the data obtained by these methods allows for modification of the treatment plan during treatment, which allows the optimal amount of LY582563 to be administered and the duration of treatment to be determined. Therefore, the treatment plan / medication schedule has been reasonably modified over the course of treatment, thereby administering the lowest amount of LY582563 that demonstrates satisfactory antiviral efficacy, and continued for as long as necessary to successfully infect Can be treated.

  In the present invention, the terms “recover”, “treat”, “treatment”, “therapeutic use”, or “treatment plan” are used herein to refer to prophylactic, palliative and therapeutic administration of LY582563. Include modalities to improve signs, disease states, conditions, symptoms, or disorders caused by hepatitis B virus, or prevent progression of signs, symptoms, conditions, or disorders associated with HBV infection Includes any and all uses of this purine to prevent, retard, or reverse. Accordingly, any prevention, amelioration, alleviation, reversal, or complete elimination of an unwanted sign, disease state, symptom, condition, or disorder associated with HBV infection is encompassed by the present invention. As used herein, the term “dosage unit” means, but is not limited to, an individual delivery carrier, such as a tablet or capsule, for administering a dose of active LY582563 purine.

  The following examples are provided to illustrate various aspects of the present invention and should not be construed as limiting the invention in any way.

Example 1
Multiple dose escalation study in healthy subjects and patients with chronic hepatitis B infection
A randomized, single period, multiple dose escalation, single blind study of LY582563 was conducted in two parts. Part 1 was performed on healthy subjects, and Part 2 was performed on patients with compensatory chronic HBV infection (hereinafter “patients”). This study was designed to evaluate the safety and pharmacodynamics of multiple doses of LY582563. This was the first multi-dose study conducted in humans.

  In healthy subjects who received the BID regimen for 2 weeks, the dose was increased in small increments. Safety data was reviewed before each dose increase.

Part 1 (Healthy subjects)
In Part 1, there were 4 groups, each group containing 8 subjects (6 subjects for study drug and 2 subjects for placebo). Table 1 shows the characteristics of the subjects in this study. The dosage of LY582563 was 2.5 mgBID, 5 mgBID, 10 mgBID and 15 mgBID. Except for the last day of QD administration, about 24 hours after the single administration of LY582563, multiple BID administrations of LY582563 were performed for about 14 days. Dose increases were made in each successive group of subjects. Safety and other data were reviewed as appropriate prior to each dose escalation step. Effective LC / MS / MS methods (Advion BioSciences, Inc., Ithaca, NY, USA) allowed plasma concentrations of LY582563 and its metabolites (602074, 602075 and 602076) to be up to 24 after the first and final dose of LY582563. Time was measured. Subjects were followed up at regular intervals of about 4 weeks after the last dose of LY582563.

Part 2 (patient)
In Part 2, there were 7 groups, each group containing 8-12 patients. Table 2 shows the characteristics of the subjects in this study. The dosage of LY582563 is 2.5 mgBID (n = 8), 5 mgBID (n = 8), 10 mgBID (n = 8), 2.5 mgQD (n = 8), 5 mgQD (n = 11), 10 mgQD (n = 12) and 20 mg QD (n = 11). Except for the last day of dosing (Day 29), which was QD, approximately 24 hours after a single dose of LY582563 (Day 1), multiple BID doses of LY582563 for about 28 days (i.e. Day 2 to Day 29) )went. Patients were tested only after reviewing safety data from healthy subjects of the same dose in Part 1. Patients were followed up at regular intervals of about 12 weeks after the last dose of LY582563. Plasma concentrations of LY582563 and its metabolites (602074, 602075 and 602076) were sampled for up to 24 hours after a single and final dose of LY582563. Subjects were followed up at regular intervals of about 12 weeks after the last dose of LY582563.

Treatment
LY582563 and the corresponding placebo were supplied as either 2.5 mg, 10 mg, or 20 mg tablets. LY582563 and the corresponding placebo tablets were supplied by Mitsubishi Pharma Corporation, Tokyo, Japan.
The LY582563 and placebo formulations for oral administration are as follows.

Production method
Mix LY582563, D-mannitol and corn starch. Hydroxypropyl methylcellulose dissolved in purified water is added and granulated with a fluid bed granulator. After drying, the granules are screened by a tornado mill. The screened granules are mixed with magnesium stearate and low substituted hydroxypropylcellulose by a V-type mixer. The mixed granules are compressed into tablets. Tablets are film coated with hydroxypropylmethyl-cellulose 2910, propylene glycol, titanium dioxide, and talc by conventional methods. Finally, the tablets are coated with a small amount of hardened oil.

dosage
The 2.5 mg dose was administered as 2.5 mg LY582563 (1 tablet) or the corresponding placebo (1 tablet). The 5 mg dose was administered as 2.5 mg LY582563 (2 tablets) or corresponding placebo (2 tablets). The 10 mg dose was administered as 10 mg LY582563 (1 tablet) or the corresponding placebo (1 tablet). The 15 mg dose was administered as 2.5 mg LY582563 (2 tablets) and 10 mg LY582563 (1 tablet) or placebo corresponding to the corresponding 2.5 and 10 mg LY582563 tablets. The 20 mg dose was administered as 20 mg LY582563 (1 tablet) or the corresponding placebo (1 tablet).

  Each participant was randomly assigned to each dose group and assigned to take either LY582563 or placebo. Part 1 healthy subjects and part 2 patients were allocated according to 4 randomized blocks (placebo 1 versus study 3). The test drug was administered BID to healthy subjects in Part 1 (Table 7) and BID or QD was administered to patients in Part 2 (Table 8).

^* Dose increases were made in each successive group. Safety and other data were reviewed as appropriate prior to each dose escalation step.

^* The order of grouping was based on the time the group started study drug therapy. The patient was tested only after reviewing safety data from healthy subjects who were at the same or equivalent dose levels in Part 1. In this study, QD dosing for patients began with 2.5 mg (3A and 3B) followed by 10 mg (3B and 4B) and 20 mg (4A). After reviewing the safety data of healthy subjects at 15 mg BID dose, it was decided not to test for QD doses higher than 20 mg. Therefore, a 5 mg QD dose was tested in the 5A group.
^** One subject in the 5A dose group randomly assigned to receive LY582563 discontinued the study one day prior to study drug administration.

Pharmacodynamic test
Blood samples were collected from patients infected with hepatitis B, and the plasma concentration of HBV / DNA amount in Part 2 of the study was measured. The amount of plasma HBV DNA was measured using an effective HBV polymerase chain reaction (PCR) assay (NGI HBV SuperQuant ™ quantitative PCR assay; National Genetics Institute, Los Angeles, CA, USA). All HBV / DNA amounts were converted to a log ₁₀ scale. The pre-dose measurement was defined as the geometric mean of the non-log transformed data obtained before dosing. The maximum log change from the pre-dose value of the HBV / DNA value in the subject during the treatment period was derived by subtracting the minimum HBV / DNA value during the treatment period from the pre-dose value. In 2 patients (2108 and 3308, both relative to placebo treatment), the maximum log change was assigned a value of zero for 2 patients whose treatment HBV DNA values were all greater than pre-dose values.

  Calculate the area under the curve (AUC) for the change in logarithm between each HBV / DNA time curve and the horizontal line through the pre-dose score to take into account the rate of decrease and recovery of HBV / DNA values during the study period did. If it is higher than the pre-dose score, first convert the HBV / DNA value to the pre-dose score, calculate the area between the curve and the pre-dose score horizontal line using the trapezoidal method, The difference in spacing was adjusted.

In the analysis, 3 HBV / DNA values that are higher than the specified test range (5 × 10 ⁹ genome copies / mL) without performing any dilution assay are assigned and assigned a value of 5 × 10 ⁹ copies / mL. One HBV DNA value below the tested range (100 genome copies / mL) was assigned a value of 100 copies / mL.

Plasma samples for HBV DNA are available from the National Genetics Institute (Los Angeles, Calif., USA) using a certified NGI HBV SuperQuantTM quantitative PCR assay with 100 genome copies / mL to 5 × 10 ⁹ genomes. Quantified with the specified assay range of copies / mL. Approximately 5 mL of blood sample is collected by screening approximately 4 and 2 weeks before and immediately prior to dosing, then dosing period (approximately 3 days after initial dosing, then 1, 2, 3, and 4 weeks) and course Measurements were taken during the observation period (5, 6, 8, 12, and 16 weeks after the first dose). Depending on the level of hepatitis B virus suppression 8 weeks after dosing, additional samples will be taken from the subject approximately 10 and 14 weeks after the first dose.

  On the dosing day, blood samples are collected and administered early in the morning. Sample collection times were adjusted, added, or deleted based on actual issues, clinical needs, or need for additional pharmacodynamic data.

  Mean and standard deviation of HBV DNA values were obtained at each scheduled time point for each dosing group.

Regression models were used to compare the average parameter values of the dosing groups with respect to log reduction, maximum log change and AUC for log change on day 29. The dose level incorporated within the dosing schedule was used as a fixed effect and the pre-dose value was used as a covariate. Differences in mean values were obtained with 95% confidence intervals. The exploratory E _max model (J. Gabrielsson and D. Weiner (1998), Pharmacokinetic and Pharmacodynamic Data Analysis: Concepts and Applications, 2nd edition, Swedish Pharmaceutical Press) is also applied to the above parameters, and the total daily dose is an explanatory variable. A well-matched regression curve was obtained, including as

Pharmacodynamic evaluation
The characteristics of the average logarithmic change of HBV • DNA are shown in FIGS. The placebo group consistently showed a mean value of zero on each measurement day (ie, not significantly different from the pre-dose value). In the case of BID dosing, the 2.5, 5, and 10 mg doses of LY582563 linearly decreased the HBV · DNA log scale during the treatment period (FIG. 1). Similar observations were made in the QD dosing group, but the rate of decline appeared to be slower in the 2.5 mg dose group, and the 5 mg group showed a slower rate of decline after day 7 (FIG. 2). In general, the HBV / DNA value gradually recovered to the pre-dose value after the final administration of LY582563, and the recovery time to the pre-dose value or the 1/2 log change of the pre-dose value increased at higher doses. Seem. Mean HBV DNA in the 10 mg BID, 10 mg QD, and 20 mg QD groups is low compared to pre-dose values on day 112 (Figures 1 and 2), suggesting that viral load does not recover to pre-dose values after a 3-month period doing.

  A distribution diagram of logarithmic change from the pre-administration value of HBV / DNA is shown in FIG. A pairwise comparison of mean log change on day 29 within each dosing schedule is listed in Table 9. Differences between pairs were tested at 5% level of significance without adjusting for multiplicity.

Note: Confidence intervals that do not include zero (shown in parentheses) indicate statistically significant differences between the two treatments at the 5% level.
^* One subject had no HBV DNA data for day 29.

  For BID dosing, the mean log change at day 29 appeared to be 2.02, 1.98, and 2.53 for the 2.5, 5, and 10 mg dose groups, respectively. All mean log changes were statistically significant when compared to those in the placebo group. There was no statistically significant difference between the three BID dose levels. For QD dosing, the mean log change observed on day 29 was 1.52, 1.96, 2.49, and 2.50 for the 2.5, 5, 10, and 20 mg dose groups, respectively. All mean log changes were statistically significant when compared to those in the placebo group. The mean log change in the 10 and 20 mg dose groups was similar and significantly different from that in the 2.5 mg group.

The E _max model including the total daily dose as an explanatory variable was applied to the logarithmic change (Fig. 4) to facilitate the understanding of the relationship between the pharmacodynamic effect and the total daily dose. The matched regression curve suggests that a plateau was obtained for HBV / DNA reduction at a total daily dose of about 10 mg.

  A pairwise comparison of the mean maximum log change in HBV DNA values during the treatment period within each dosing schedule is listed in Table 10. Pair differences were tested at the 5% level of significance without any multiple adjustment.

  Note: Confidence intervals that do not include zero (shown in parentheses) indicate statistically significant differences between the two treatments at the 5% level.

  For BID dosing, the mean maximum log change over the treatment period appeared to be 2.39, 1.82, and 2.93 for the 2.5, 5, and 10 mg dose groups, respectively. All mean maximum log changes were statistically significant when compared to that of the placebo group. There appeared to be a significant difference in mean log change between the 10 mg and 5 mg dose groups, but there was no significant difference between the 10 mg and 2.5 mg dose groups. For QD dosing, the mean maximum log change observed during the treatment period was 1.53, 2.19, 2.58, and 2.68 for the 2.5, 5, 10, and 20 mg dose groups, respectively. All mean log reductions were statistically significant when compared to those in the placebo group. The mean log change in the 5, 10, and 20 mg dose groups appeared similar and was significantly different from the 2.5 mg group.

  At 10 mg BID, 10 mg QD, and 20 mg QD, all patients showed a maximum log change in HBV DNA greater than 1.5 during the treatment period.

An E _max model with total daily dose as an explanatory variable was fitted to the maximum log change (FIG. 5). The matched regression curve suggests that a plateau was obtained for maximum HBV / DNA reduction during the treatment period at a total daily dose of about 10 mg.

To further clarify, FIG. 4 shows the logarithmic change of HBV DNA on day 29 along with a matched E _max model based on total daily dose (TDD), and FIG. 5 is based on TDD. Together with the matched E _max model, shows the maximum log change in HBV DNA over the treatment period. In some patients, the maximum log change occurred during a period other than day 29, so both figures should be taken into account. Considering these figures together, we can better understand the pharmacodynamic effects on dose, that is, the characteristics of HBV and DNA changes.

  Within each dosing schedule, a pairwise comparison of the average change in AUC from the first dose to the end of the study period (day 112) is listed in Table 11. Pair differences were tested at the 5% level of significance without any multiple adjustment.

  Note: Confidence intervals that do not include zero (shown in parentheses) indicate statistically significant differences between the two treatments at the 5% level.

  For BID dosing, the mean AUC for log change after the first dose in the 2.5, 5, and 10 mg dose groups appeared to be statistically significant when compared to that in the placebo group. There was also a significant difference in mean AUC for log changes between the 10 mg and 2.5 mg dose groups. For QD dosing, the average AUC observed for log changes in the 2.5, 5, 10, and 20 mg dose groups was statistically significant compared to that of the placebo group. The mean AUC for log change in the 10 and 20 mg dose groups appeared similar and was significantly different from that in the 2.5 mg group. There was also a trend of significant difference between 10 mg and 5 mg doses.

An E _max model with total daily dose as an explanatory variable was applied to the AUC for log change, which was similar to the E _max model for log change of HBV • DNA (FIG. 6). The consistent regression curve suggests that a total daily dose of 10 mg provides 80% of the effect of 20 mg total daily dose on the AUC log change of HBV DNA.

Conclusions on the efficacy of LY582563 As demonstrated by the data presented above, all doses of LY582563 tested in this study were accompanied by a decrease in plasma HBV DNA during the treatment period, which was dose-dependent It seemed to be. The 10 mg QD, 10 mg BID, and 20 mg QD dose groups had a mean decrease in HBV DNA on day 29 (Figure 3), mean maximum log virus reduction (Table 10), and mean AUC change in log virus reduction after the first dose ( As seen in Table 11), maximum virus suppression was shown. This reduced level obtained with drugs such as interferon alpha, lamivudine, and adefovir has been shown to continue the progression of HBV / DNA suppression, and the continued increase in seroconversion with continued treatment . This data suggests a plateau of effect before a total daily dose of 10 mg. A dosage of about 5 mg per day to about 10 mg per day, or about 7.5 mg per day to about 10 mg per day may be optimal. In each case, the upper limit of these ranges may extend to about 12 or about 12.5 mg per day. These dosages can be administered in the form of tablets or capsules.

  The viral load gradually returned to pre-dose values after cessation of LY582563 treatment, and recovery of viral load was slower in higher dose groups (FIGS. 1 and 2). The slower recovery of virus replication at higher dose groups may be due to greater suppression of the virus during treatment and smaller residual portions of infected hepatocytes. The virus dose in each dose group on day 29 is different because the virus dose in the higher dose group is lower (Figures 1 and 2) compared to the higher and lower dose groups. Therefore, after cessation of LY582563 treatment, no analysis was performed to determine recovery of viral load relative to pre-dose values. This study was not designed to detect the mean log change between dose groups from pre-dose HBV DNA values on day 29.

Safety of LY582563 The most commonly reported side effects in healthy subjects were sleepiness, epigastric pain, and diarrhea. In patients with chronic hepatitis B infection, the most commonly reported side effects were sleepiness, headache, and intravenous cannula-related events. Since the incidence of side effects at the highest dose level (10 mg BID, 20 mg QD) was similar to or lower than placebo, the dose did not seem to be related to side effects. None of the subjects discontinued the study due to side effects.

  Both healthy subjects (about 30%) and compensatory chronic HBV-infected patients (about 50-70%) have the highest incidence of ALT increase from pre-dose values in higher dose groups, leading to ALT increase The dose seemed to be related. The maximum range of ALT increase by LY582563 was about 3 times the upper limit of the standard.

From the assessment of clinical biochemical markers (serum phosphorus, urea, and creatinine) and clinical urine markers (LDH / creatinine and β-NAG / creatinine ratio) in both healthy subjects and patients, dose-related renal urine Tubular toxicity was not apparent. Laboratory results of plasma lactate and serum creatine kinase also suggested that there was no dose-related mitochondrial or musculoskeletal toxicity after repeated dosing with the tested LY582563 dose. None of the healthy subjects or patients receiving LY582563 had vital signs, electrocardiogram (ECG) clinically significant abnormalities, and prolongation of QT _c longer than 30 ms.

The results shown above are for patients with compensatory chronic HBV infection when administered to healthy subjects in multiple doses of 2.5 mg to 30 mg daily for up to 14 days, and for up to 28 days with multiple doses of 2.5 mg to 20 mg daily LY582563 has been shown to be safe and well tolerated. There appeared to be a dose-related relationship to ALT elevation, as the highest incidence of ALT elevation from pre-dose in the higher dose group in both healthy subjects and patients with compensatory chronic HBV infection. The maximum range of ALT rise attributed to LY582563 was about three times the upper limit of the standard. Other biochemical markers showed no relationship to dose. There were no significant abnormal signs of survival or ECG (including assessment of QT _c prolongation) in healthy subjects and patients with compensated chronic HBV infection. All doses of LY582563 were associated with reduced viral load (plasma HBV / DNA) that appeared to be dose related during the treatment period. The greatest decrease was seen in the 10 mg QD, 10 mg BID, and 20 mg QD dosage groups, resulting in an average log ₁₀ decrease of about 2.5 after 4 weeks of treatment. The viral load gradually recovered to the pre-dose value after cessation of LY582563 treatment. This appeared to be delayed at higher doses. Based on these results, a suitable LY582563 dose that balances safety and efficacy is about 2.5 mg to about 20 mg per patient per day. These dosages may have to be administered to patients over a period of days, weeks, months or years to remedy or control the consequences of an unfavorable condition of HBV infection.

  While the invention has been described above, it will be appreciated that the invention may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all modifications apparent to those skilled in the art are intended to be included within the scope of the following claims. .

FIG. 1 shows the characteristics of the mean log change of HBV • DNA together with the standard deviation bar (BID group). FIG. 2 shows the characteristics of the mean log change of HBV • DNA together with the standard deviation bar (QD group). FIG. 3 is a distribution diagram showing the logarithmic change of HBV · DNA on the 29th day together with the mean value and the standard deviation bar. Individual open symbols, i.e., squares, triangles, etc., represent individual patient data points. These are different to show some differences between the columns of data at different daily doses, but this is not always necessary because the columns are clearly distinguished. The filled symbols in each column represent the average value. The outer boundary of the bar represents the standard deviation from its mean value. FIG. 4 shows the log change of HBV • DNA on day 29 along with a congruent E _max model based on total daily dose (TDD). FIG. 5 shows the maximum log change of HBV • DNA during the treatment period, along with a matched E _max model based on total daily dose (TDD). FIG. 6 shows the area under the curve (AUC) versus log change in HBV • DNA after the first dose, along with a matched E _max model based on the total daily dose (TDD).

Claims (16)

  About 2.5 mg to about 20 mg of 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine, per dosage unit; And a pharmaceutical composition for oral administration in dosage unit form, comprising a pharmaceutically acceptable carrier, diluent or excipient   The composition of claim 1 in the form of a tablet or capsule.   The 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is about 2.5 mg per dosage unit, about 3. A composition according to claim 1 or 2 present in an amount of 5 mg, about 10 mg, about 15 mg, or about 20 mg.   2-Amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] for the preparation of a medicament for treating human patients suffering from hepatitis B virus infection -6- (4-methoxyphenylthio) purine, wherein the medicament is prepared for oral administration, and the medicament is in dosage unit form, wherein about 2.5 mg to about 20 mg of the purine is administered per dosage unit. Including use.   Use according to claim 4, wherein the medicament is in the form of a tablet or capsule.   The 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is about 2.5 mg per dosage unit, about 6. Use according to claim 4 or 5, present in an amount of 5 mg, about 10 mg, about 15 mg, or about 20 mg.   A method of treating a human patient suffering from a hepatitis B virus infection, said patient comprising 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] Administering ethyl] -6- (4-methoxyphenylthio) purine in a total amount of about 2.5 mg to about 20 mg of the purine per day.   The 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is added before the purine is administered. 8. The method of claim 7, wherein the method is administered to the patient for a period of time sufficient to reduce the patient's plasma HBV / DNA level relative to the patient's plasma HBV / DNA level. 9. The method of claim 8, wherein the plasma HBV / DNA level of the patient is reduced to at least about 10 ⁴ copies / mL compared to the patient's plasma HBV / DNA level prior to administration of the purine.   The 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is the hepatitis B virus of the patient. 8. The method of claim 7, wherein the method is administered to the patient for a period of time sufficient to ameliorate a sign, symptom, condition, or disorder caused by.   11. The method of claim 10, wherein the sign, symptom, condition, or disorder is selected from the group consisting of liver fibrosis, cirrhosis, inflammatory liver disease, and liver cancer.   The 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is a pharmaceutically acceptable oral composition. 12. The method according to any one of claims 7 to 11, wherein the method is administered to the patient in the form of a product.   13. The method according to any one of claims 7 to 12, wherein the pharmaceutically acceptable oral composition is in the form of a tablet or capsule.   14. A method according to any one of claims 8 to 13, wherein the period is days, weeks, months or years.   The 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is about 2.5 mg per day to the patient. 15. The method of any one of claims 7 to 14, wherein the dose is administered in a total amount of about 5 mg, about 10 mg, about 15 mg, or about 20 mg.   The total amount of the 2-amino-9- [2- [bis (2,2,2-trifluoroethoxy) phosphonylmethoxy] ethyl] -6- (4-methoxyphenylthio) purine is a single dose. 16. A method according to any one of claims 7 to 15, wherein the method is administered in divided doses, or in total, the total amount per day.

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