U.S. patent application number 16/040959 was filed with the patent office on 2019-04-18 for combination formulation of two antiviral compounds.
The applicant listed for this patent is Gilead Pharmasset LLC. Invention is credited to Ben Chal, Erik Mogalian, Reza Oliyai, Rowchanak Pakdaman, Dimitrios Stefanidis, Vahid Zia.
Application Number | 20190111068 16/040959 |
Document ID | / |
Family ID | 50102269 |
Filed Date | 2019-04-18 |
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United States Patent
Application |
20190111068 |
Kind Code |
A1 |
Chal; Ben ; et al. |
April 18, 2019 |
COMBINATION FORMULATION OF TWO ANTIVIRAL COMPOUNDS
Abstract
Disclosed are pharmaceutical compositions having an effective
amount of substantially amorphous ledipasvir and an effective
amount of substantially crystalline sofosbuvir.
Inventors: |
Chal; Ben; (Millbrae,
CA) ; Mogalian; Erik; (San Francisco, CA) ;
Oliyai; Reza; (Burlingame, CA) ; Pakdaman;
Rowchanak; (San Carlos, CA) ; Stefanidis;
Dimitrios; (Mountain View, CA) ; Zia; Vahid;
(San Carlos, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Gilead Pharmasset LLC |
Foster City |
CA |
US |
|
|
Family ID: |
50102269 |
Appl. No.: |
16/040959 |
Filed: |
July 20, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15393847 |
Dec 29, 2016 |
10039779 |
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16040959 |
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14868062 |
Sep 28, 2015 |
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15393847 |
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14168264 |
Jan 30, 2014 |
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14868062 |
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61907332 |
Nov 21, 2013 |
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61897793 |
Oct 30, 2013 |
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61870729 |
Aug 27, 2013 |
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61828899 |
May 30, 2013 |
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61772292 |
Mar 4, 2013 |
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61759320 |
Jan 31, 2013 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/1623 20130101;
A61K 31/513 20130101; A61P 31/14 20180101; A61K 31/4025 20130101;
A61K 9/2013 20130101; A61K 31/4985 20130101; A61K 31/497 20130101;
A61K 9/28 20130101; A61K 31/5377 20130101; A61K 31/7072 20130101;
A61K 31/381 20130101; A61K 31/7076 20130101; A61K 31/5025 20130101;
A61K 9/2018 20130101; A61K 9/2009 20130101; A61P 31/12 20180101;
A61K 31/675 20130101; A61K 31/4709 20130101; A61K 9/2027 20130101;
A61P 1/16 20180101; A61K 9/1635 20130101; A61K 31/439 20130101;
A61K 31/7056 20130101; A61K 31/4184 20130101; A61K 9/2054 20130101;
A61K 31/4178 20130101; A61K 9/209 20130101; A61K 31/4184 20130101;
A61K 2300/00 20130101; A61K 31/7072 20130101; A61K 2300/00
20130101; A61K 31/7056 20130101; A61K 2300/00 20130101; A61K
31/4709 20130101; A61K 2300/00 20130101; A61K 31/5025 20130101;
A61K 2300/00 20130101; A61K 31/4025 20130101; A61K 2300/00
20130101; A61K 31/381 20130101; A61K 2300/00 20130101; A61K 31/7076
20130101; A61K 2300/00 20130101; A61K 31/513 20130101; A61K 2300/00
20130101; A61K 31/4985 20130101; A61K 2300/00 20130101; A61K 31/675
20130101; A61K 2300/00 20130101; A61K 31/497 20130101; A61K 2300/00
20130101; A61K 31/5377 20130101; A61K 2300/00 20130101; A61K 31/439
20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/7072 20060101
A61K031/7072; A61K 31/4184 20060101 A61K031/4184; A61K 9/20
20060101 A61K009/20; A61K 31/7056 20060101 A61K031/7056; A61K
31/439 20060101 A61K031/439; A61K 9/28 20060101 A61K009/28; A61K
31/7076 20060101 A61K031/7076; A61K 31/675 20060101 A61K031/675;
A61K 31/5377 20060101 A61K031/5377; A61K 31/5025 20060101
A61K031/5025; A61K 31/4985 20060101 A61K031/4985; A61K 31/497
20060101 A61K031/497; A61K 31/4025 20060101 A61K031/4025; A61K
31/381 20060101 A61K031/381; A61K 31/513 20060101 A61K031/513; A61K
9/16 20060101 A61K009/16; A61K 9/24 20060101 A61K009/24; A61K
31/4178 20060101 A61K031/4178; A61K 31/4709 20060101
A61K031/4709 |
Claims
1. A pharmaceutical composition comprising: a) an effective amount
of ledipasvir having the formula: ##STR00017## wherein the
ledipasvir is substantially amorphous; and b) an effective amount
of sofosbuvir having the formula: ##STR00018## wherein the
sofosbuvir is substantially crystalline.
2. The pharmaceutical composition of claim 1, wherein ledipasvir is
formulated as a solid dispersion comprising ledipasvir dispersed
within a polymer matrix formed by a pharmaceutically acceptable
polymer.
3. The pharmaceutical composition of claim 2, wherein the polymer
is copovidone.
4. The pharmaceutical composition of claim 3, wherein the weight
ratio of ledipasvir to copovidone in the solid dispersion is about
1:1.
5. The pharmaceutical composition of claim 3, comprising a) about
40% w/w of sofosbuvir and b) about 18% w/w of the solid dispersion
comprising ledipasvir.
6. The pharmaceutical composition of claim 5, further comprising a)
about 5 to about 25% w/w lactose monohydrate, b) about 5 to about
25% w/w microcrystalline cellulose, c) about 1 to about 10% w/w
croscarmellose sodium, d) about 0.5 to about 3% w/w colloidal
silicon dioxide, and e) about 0.1 to about 3% w/w magnesium
stearate.
7. A pharmaceutical dosage form comprising the pharmaceutical
composition of claim 1, comprising about 90 mg of ledipasvir and
about 400 mg of sofosbuvir.
8. The pharmaceutical dosage form of claim 7, wherein the
ledipasvir is formulated as a solid dispersion within a polymer
matrix of copovidone.
9. The pharmaceutical dosage form of claim 8, wherein the amount of
copovidone is about 90 mg.
10. The pharmaceutical dosage form of claim 9, further comprising:
(a) about 165 mg of lactose monohydrate; (b) about 180 mg of
microcrystalline cellulose; (c) about 50 mg of croscarmellose
sodium; (d) about 10 mg of colloidal silicon dioxide; and (e) about
15 mg of magnesium stearate.
11. The pharmaceutical dosage form of claim 9 which is in the form
of a tablet comprising a film coating.
12. A method of treating a patient infected with hepatitis C virus
comprising administering to the patient a therapeutically effective
amount of a pharmaceutical composition of claim 1.
13. The method of claim 12, wherein the pharmaceutical composition
is administered for about 24 weeks or less.
14. The method of claim 12, wherein the pharmaceutical composition
is administered for about 12 weeks or less.
15. The method of claim 12, wherein the pharmaceutical composition
is administered for about 8 weeks or less.
16. The method of claim 12, wherein the pharmaceutical composition
is administered for about 6 weeks or less.
17. The method of claim 12, wherein the pharmaceutical composition
is administered once daily for about 12 weeks or less and wherein
the hepatitis C virus is genotype 1, 2, 3, 4, 5, or 6.
18. The method of claim 12, wherein the pharmaceutical composition
is administered once daily for about 8 weeks or less and wherein
the hepatitis C virus is genotype 1, 2, 3, 4, 5, or 6.
19. The method of claim 12, wherein the pharmaceutical composition
is administered once daily for about 6 weeks or less and wherein
the hepatitis C virus is genotype 1, 2, 3, 4, 5, or 6.
20. The method of claim 17, wherein the hepatitis C virus is
genotype 1a or 1b.
21. The method of claim 18, wherein the hepatitis C virus is
genotype 1a or 1b.
22. The method of claim 19, wherein the hepatitis C virus is
genotype 1a or 1b.
23. The method of claim 12, wherein the pharmaceutical composition
is administered once daily for about 12 weeks and wherein the
hepatitis C virus is genotype 1a, 1b, 2a, 2b, 2c, 2d, 3a, 3b, 3c,
3d, 3e, 3f, 4a, 4b, 4c, 4d, 4e, 4f, 4g, 4h, 4i, 5a, or 6a.
24. The method of claim 12, wherein the pharmaceutical composition
is administered once daily for about 8 weeks and wherein the
hepatitis C virus is genotype 1a, 1b, 2a, 2b, 2c, 2d, 3a, 3b, 3c,
3d, 3e, 3f, 4a, 4b, 4c, 4d, 4e, 4f, 4g, 4h, 4i, 5a, or 6a.
25. The method of claim 12, further comprising administering
ribavirin.
26. The method of claim 12, wherein the treatment does not include
interferon.
27. The method of claim 12, wherein the treatment does not include
ribavirin.
28. The method of claim 12, wherein the treatment does not include
interferon or ribavirin.
29. The method of claim 12, further comprising administering an NS3
protease inhibitor.
30. The method of claim 12, further comprising administering
simeprevir.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 15/393,847, filed Dec. 29, 2016, now U.S. Pat. No. 10,039,779,
which is a continuation of U.S. application Ser. No. 14/868,062,
filed Sep. 28, 2015, now abandoned, which is a continuation of U.S.
application Ser. No. 14/168,264, filed Jan. 30, 2014, now
abandoned, which claims the benefit under 35 U.S.C. .sctn. 119(e)
to U.S. Provisional Application No. 61/759,320, filed on Jan. 31,
2013, U.S. Provisional Application No. 61/772,292, filed on Mar. 4,
2013, U.S. Provisional Application No. 61/828,899, filed on May 30,
2013, U.S. Provisional Application No. 61/870,729, filed on Aug.
27, 2013, U.S. Provisional Application No. 61/897,793, filed on
Oct. 30, 2013, and U.S. Provisional Application No. 61/907,332,
filed on Nov. 21, 2013, the entirety of which are all incorporated
herein by reference.
BACKGROUND
[0002] Hepatitis C is recognized as a chronic viral disease of the
liver which is characterized by liver disease. Although drugs
targeting the liver are in wide use and have shown effectiveness,
toxicity and other side effects have limited their usefulness.
Inhibitors of hepatitis C virus (HCV) are useful to limit the
establishment and progression of infection by HCV as well as in
diagnostic assays for HCV.
[0003] Ledipasvir is a selective inhibitor of non-structural
protein 5A (NS5A), which has been described previously (see, for
example, WO 2010/132601). The chemical name of ledipasvir is
(1-{3-[6-(9,9-difluoro-7-{2-[5-(2-methoxycarbonylamino-3-methyl-butyryl)--
5-aza-spiro[2.4]hept-6-yl]-3H-imidazol-4-yl}-9H-fluoren-2-yl)-1H-benzoimid-
azol-2-yl]-2-aza-bicyclo[2.2.1]heptane-2-carbonyl}-2-methyl-propyl)-carbam-
ic acid methyl ester.
[0004] Sofosbuvir (SOF) is a selective inhibitor of non-structural
protein 5B (NS5B) (see, for example, WO 2010/132601 and U.S. Pat.
No. 7,964,580). The chemical name of sofosbuvir is (S)-isopropyl
2-(((S)-(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-flu-
oro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)ami-
no) propanoate.
SUMMARY
[0005] The present disclosure provides, in some embodiments, a
pharmaceutical composition comprising ledipasvir in a substantially
amorphous form and sofosbuvir in a substantially crystalline
form.
[0006] Ledipasvir has the chemical name of
(1-{3-[6-(9,9-difluoro-7-{2-[5-(2-methoxycarbonylamino-3-methyl-butyryl)--
5-aza-spiro[2.4]hept-6-yl]-3H-imidazol-4-yl}-9H-fluoren-2-yl)-1H-benzoimid-
azol-2-yl]-2-aza-bicyclo[2.2.1]heptane-2-carbonyl}-2-methyl-propyl)-carbam-
ic acid methyl ester, and has the following chemical formula:
##STR00001##
[0007] Sofosbuvir (SOF) has the chemical name of (S)-isopropyl
2-(((S)-(((2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-flu-
oro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)ami-
no)propanoate and has the following chemical formula:
##STR00002##
[0008] In some embodiments, provided is a pharmaceutical
composition comprising: a) an effective amount of ledipasvir,
wherein ledipasvir is substantially amorphous; and b) an effective
amount of sofosbuvir wherein sofosbuvir is substantially
crystalline.
[0009] Further embodiments of the disclosure relate to
pharmaceutical dosage forms and tablets. The disclosure also
provides methods for using the combination in the treatment of
hepatitis C.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] FIG. 1 is a XRPD pattern of the solid dispersion formulation
of ledipasvir comprising copovidone in a drug:polymer ratio of 1:1.
As shown by the XRPD, the solid dispersion is in the amorphous
state.
[0011] FIG. 2 is a modulated differential scanning calorimetry
(DSC) curve of the solid dispersion of ledipasvir comprising
copovidone in a drug:polymer ratio of 1:1. The glass transition
temperature of the solid dispersion is about 140.degree. C.
[0012] FIG. 3 shows a solid state characterization of the solid
dispersion formulation of ledipasvir comprising copovidone in a
drug:polymer ratio of 1:1 by solid state nuclear magnetic resonance
(SS-NMR).
[0013] FIG. 4 is a Fourier-transformed Raman spectra of the solid
dispersion of ledipasvir comprising copovidone in a drug:polymer
ratio of 1:1.
[0014] FIG. 5 shows the dissolution of sofosbuvir in the sofosbuvir
(400 mg)/ledipasvir (90 mg) combination described in Example 7.
[0015] FIG. 6 shows the dissolution of ledipasvir in the sofosbuvir
(400 mg)/ledipasvir (90 mg) combination formulation described in
Example 3.
[0016] FIG. 7, with panels A-D, shows the HCV RNA levels during 12
weeks of treatment and 24 weeks post-treatment for treatment naive
(A) or null responder (B) patients treated with sofosbuvir (SOF)
and ribavirin (RBV) and for treatment naive (C) or null responder
(D) patients treated with sofosbuvir (SOF), ledipasvir and
ribavirin (RBV). This data and experimental method are further
described in Example 5.
[0017] FIG. 8, with panels A and B, presents charts to show that
all three formulations had comparable dissolution performance,
similar to that of the single-agent controls. This is more
thoroughly described in Example 7.
[0018] FIG. 9 presents the pH-solubility profile of ledipasvir at
room temperature (RT). The line is the nonlinear least-square
regression fit using equation
S.sub.T=S.sub.0[(1+10.sup.(pKa1-pH)+10.sup.(pKa1+pKa2-2pH))] with
an intrinsic solubility (S.sub.0) of 0.04 .mu.g/mL and a weakly
basic pKa1 and pKa2 values of 5.0 and 4.0, respectively. This is
more thoroughly described in Example 8.
[0019] FIG. 10 shows the study design for treatment naive
(non-cirrhotic) and for null responders (50% cirrhotic) for
patients treated with a fixed dose combination of sofosbuvir (SOF)
and ledipasvir, with and without ribavirin (RBV) for 8 and 12
weeks. The data and experimental method are described in Example
9.
[0020] FIG. 11 shows the results for treatment naive
(non-cirrhotic) and for null responders (50% cirrhotic) for
patients treated with a fixed dose combination of sofosbuvir (SOF)
and ledipasvir, with and without ribavirin (RBV) for 8 and 12
weeks. This data and experimental method are further described in
Example 9.
DETAILED DESCRIPTION
1. Definitions
[0021] As used in the present specification, the following words
and phrases are generally intended to have the meanings as set
forth below, except to the extent that the context in which they
are used indicates otherwise.
[0022] As used herein, the term "about" used in the context of
quantitative measurements means the indicated amount .+-.10%, or
alternatively .+-.5%, or .+-.1%. For example, with a .+-.10% range,
"about 2:8" can mean 1.8-2.2:7.2-8.8.
[0023] The term "amorphous" refers to a state in which the material
lacks long range order at the molecular level and, depending upon
temperature, may exhibit the physical properties of a solid or a
liquid. Typically such materials do not give distinctive X-ray
diffraction patterns and, while exhibiting the properties of a
solid, are more formally described as a liquid. Upon heating, a
change from solid to liquid properties occurs which is
characterized by a change of state, typically second order (glass
transition).
[0024] The term "crystalline" refers to a solid phase in which the
material has a regular ordered internal structure at the molecular
level and gives a distinctive X-ray diffraction pattern with
defined peaks. Such materials when heated sufficiently will also
exhibit the properties of a liquid, but the change from solid to
liquid is characterized by a phase change, typically first order
(melting point).
[0025] The term "substantially amorphous" as used herein is
intended to mean that greater than 70%; or greater than 75%; or
greater than 80%; or greater than 85%; or greater than 90%; or
greater than 95%, or greater than 99% of the compound present in a
composition is in amorphous form. "Substantially amorphous" can
also refer to material which has no more than about 20%
crystallinity, or no more than about 10% crystallinity, or no more
than about 5% crystallinity, or no more than about 2%
crystallinity.
[0026] The term "substantially crystalline" as used herein is
intended to mean that greater than 70%; or greater than 75%; or
greater than 80%; or greater than 85%; or greater than 90%; or
greater than 95%, or greater than 99% of the compound present in a
composition is in crystalline form. "Substantially crystalline" can
also refer to material which has no more than about 20%, or no more
than about 10%, or no more than about 5%, or no more than about 2%
in the amorphous form.
[0027] The term "polymer" refers to a chemical compound or mixture
of compounds consisting of repeating structural units created
through a process of polymerization. Suitable polymers useful in
this invention are described throughout.
[0028] The term "polymer matrix" as used herein is defined to mean
compositions comprising one or more polymers in which the active
agent is dispersed or included within the matrix.
[0029] The term "solid dispersion" refers to the dispersion of one
or more active agents in a polymer matrix at solid state prepared
by a variety of methods, including spray drying, the melting
(fusion), solvent, or the melting-solvent method.
[0030] The term "amorphous solid dispersion" as used herein, refers
to stable solid dispersions comprising an amorphous active agent
and a polymer. By "amorphous active agent," it is meant that the
amorphous solid dispersion contains active agent in a substantially
amorphous solid state form. In some aspects, as shown by the XRPD
in FIG. 1, the solid dispersion is in the amorphous state, and the
glass transition temperature of the solid dispersion is about
140.degree. C. (see FIG. 2).
[0031] The term "pharmaceutically acceptable" indicates that the
material does not have properties that would cause a reasonably
prudent medical practitioner to avoid administration of the
material to a patient, taking into consideration the disease or
conditions to be treated and the respective route of
administration. For example, it is commonly required that such a
material be essentially sterile, e.g., for injectibles.
[0032] The term "pharmaceutically acceptable polymer" refers to a
polymer that does not have properties that would cause a reasonably
prudent medical practitioner to avoid administration of the
material to a patient, taking into consideration the disease or
conditions to be treated and the respective route of
administration.
[0033] The term "carrier" refers to a glidant, diluent, adjuvant,
excipient, or vehicle etc with which the compound is administered,
without limitation. Examples of carriers are described herein and
also in "Remington's Pharmaceutical Sciences" by E. W. Martin.
[0034] The term "diluent" refers to chemical compounds that are
used to dilute the compound of interest prior to delivery. Diluents
can also serve to stabilize compounds. Non-limiting examples of
diluents include starch, saccharides, disaccharides, sucrose,
lactose, polysaccharides, cellulose, cellulose ethers,
hydroxypropyl cellulose, sugar alcohols, xylitol, sorbitol,
maltitol, microcrystalline cellulose, calcium or sodium carbonate,
lactose, lactose monohydrate, dicalcium phosphate, cellulose,
compressible sugars, dibasic calcium phosphate dehydrate, mannitol,
microcrystalline cellulose, and tribasic calcium phosphate.
[0035] The term "binder" when used herein relates to any
pharmaceutically acceptable film which can be used to bind together
the active and inert components of the carrier together to maintain
cohesive and discrete portions. Non-limiting examples of binders
include hydroxypropylcellulose, hydroxypropylmethylcellulose,
povidone, copovidone, and ethyl cellulose.
[0036] The term "disintegrant" refers to a substance which, upon
addition to a solid preparation, facilitates its break-up or
disintegration after administration and permits the release of an
active ingredient as efficiently as possible to allow for its rapid
dissolution. Non-limiting examples of disintegrants include maize
starch, sodium starch glycolate, croscarmellose sodium,
crospovidone, microcrystalline cellulose, modified corn starch,
sodium carboxymethyl starch, povidone, pregelatinized starch, and
alginic acid.
[0037] The term "lubricant" refers to an excipient which is added
to a powder blend to prevent the compacted powder mass from
sticking to the equipment during the tabletting or encapsulation
process. It aids the ejection of the tablet form the dies, and can
improve powder flow. Non-limiting examples of lubricants include
magnesium stearate, stearic acid, silica, fats, calcium stearate,
polyethylene glycol, sodium stearyl fumarate, or talc; and
solubilizers such as fatty acids including lauric acid, oleic acid,
and C.sub.8/C.sub.10 fatty acid.
[0038] The term "film coating" refers to a thin, uniform, film on
the surface of a substrate (e.g. tablet). Film coatings are
particularly useful for protecting the active ingredient from
photolytic degradation. Non-limiting examples of film coatings
include polyvinylalcohol based, hydroxyethylcellulose,
hydroxypropylmethylcellulose, sodium carboxymethylcellulose,
polyethylene glycol 4000 and cellulose acetate phthalate film
coatings.
[0039] The term "glidant" as used herein is intended to mean agents
used in tablet and capsule formulations to improve flow-properties
during tablet compression and to produce an anti-caking effect.
Non-limiting examples of glidants include colloidal silicon
dioxide, talc, fumed silica, starch, starch derivatives, and
bentonite.
[0040] The term "effective amount" refers to an amount that is
sufficient to effect treatment, as defined below, when administered
to a mammal in need of such treatment. The therapeutically
effective amount will vary depending upon the patient being
treated, the weight and age of the patient, the severity of the
disease condition, the manner of administration and the like, which
can readily be determined by one of ordinary skill in the art.
[0041] The term "treatment" or "treating," to the extent it relates
to a disease or condition includes preventing the disease or
condition from occurring, inhibiting the disease or condition,
eliminating the disease or condition, and/or relieving one or more
symptoms of the disease or condition.
[0042] The term "sustained virologic response" refers to the
absence of detectable RNA (or wherein the RNA is below the limit of
detection) of a virus (i.e. HCV) in a patient sample (i.e. blood
sample) for a specifc period of time after discontinuation of a
treatment. For example, a SVR at 4 weeks indicates that RNA was not
detected or was below the limit of dectection in the patient at 4
weeks after discontinuing HCV therapy.
[0043] The term "% w/w" as used herein refers to the weight of a
component based on the total weight of a composition comprising the
component. For example, if component A is present in an amount of
50% w/w in a 100 mg composition, component A is present in an
amount of 50 mg.
2. Pharmaceutical Compositions
[0044] The pharmaceutical compositions comprise a combination of an
effective amount of ledipasvir, wherein ledipasvir is substantially
amorphous, and an effective amount of sofosbuvir, wherein
sofosbuvir is substantially crystalline.
[0045] Such a combination composition, as the experimental examples
demonstrate, exhibit unexpected properties. Both sofosbuvir and
ledipasvir have previously been demonstrated to act as effective
anti-HCV agents. Ledipasvir, when administered alone in a
conventional formulation, however, exhibited a negative food effect
as evidenced by a roughly 2-fold decrease in exposure when given
with a high-fat meal relative to dosing in the fasted state (see,
e.g., Tables 10 and 11, Example 3). When ledipasvir is administered
in a solid dispersion formulation and in the combination with
sofosbuvir, no such negative food effect occurs (Table 12, Example
3).
[0046] In the combination composition, ledipasvir is present in a
substantially amorphous form. Compared to crystalline agents,
amorphous agents are expected to be unstable and have nonlinear
solubility and exposure profiles. The data presented herein,
however, show that ledipasvir in the combination composition is
stable under various conditions, both short-term and long-term, and
maintains high and consistent solubility and exposure profiles
(Example 6).
[0047] Further, according the conventional wisdom, it is not
advisable to co-formulate an amorphous agent with a crystalline
agent, because the crystals can serve as seeds to induce
crystallization of the amorphous agent, leading to instability of
the amorphous agent. The current data show that, however, whether
co-granulated or co-blended with sofosbuvir in the same layer or
integrated as separate layers, ledipasvir stays stable and does not
form crystals in the composition (Example 6).
[0048] It is also been discovered that, in tablet formations of the
combination composition where sofosbuvir and ledipasvir are either
co-granulated or co-blended, drug-drug interaction does not occur
(Example 7).
A. Ledipasvir
[0049] Ledipasvir has previously been described (see, for example,
WO 2010/132601) and can be prepared by methods described therein.
In one embodiment, the pharmaceutical composition comprises
ledipasvir formulated as a solid dispersion dispersed within a
polymer matrix formed by a pharmaceutically acceptable polymer. The
starting material of the solid dispersion can be a variety of forms
of ledipasvir including crystalline forms, amorphous form, salts
thereof, solvates thereof and the free base. For example, the
acetone solvate, D-tartrate salt, anhydrous crystalline free base,
amorphous free base, solvates or desolvates of ledipasvir can be
used. Solvates of ledipasvir include, for example, those described
in U.S. Publication No. 2013/0324740 (incorporated herein by
reference) such as, for example, the monoacetone solvate, diacetone
solvate, ethyl acetone solvate, isopropyl acetate solvate, methyl
acetate solvate, ethyl formate solvate, acetonitrile solvate,
tetrahydrofuran solvate, methyl ethyl ketone solvate,
tetrahydrofuran solvate, methyl ethyl ketone solvate, and methyl
tert-butyl ether solvate. Particular starting materials
contemplated to be useful are the monoacetone solvate, diacetone
solvate, anhydrous crystalline free base, D-tartrate salt,
anhydrous crystalline free base, and amorphous free base. These
forms are characterized and described in U.S. Publication No.
2013/0324496.
[0050] After dispersion with the polymer, the solid dispersion is
in the amorphous form. FIGS. 1-4 characterize the amorphous solid
dispersion comprising ledipasvir. As shown by the XRPD in FIG. 1,
the solid dispersion is in the amorphous state, and the glass
transition temperature of the solid dispersion is about 140.degree.
C.
[0051] Various techniques are well known in the art for preparing
solid dispersions including, but not limited to melt-extrusion,
spray-drying, lyophilization, and solution-evaporation.
[0052] Melt-extrusion is the process of embedding a compound in a
thermoplastic carrier. The mixture is processed at elevated
temperatures and pressures, which disperses the compound in the
matrix at a molecular level to form a solid solution. Extruded
material can be further processed into a variety of dosage forms,
including capsules, tablets and transmucosal systems.
[0053] For the solution-evaporation method, the solid dispersion
can be prepared by dissolving the compound in a suitable liquid
solvent and then incorporating the solution directly into the melt
of a polymer, which is then evaporated until a clear, solvent free
film is left. The film is further dried to constant weight.
[0054] For the lyophilization technique, the compound and carrier
can be co-dissolved in a common solvent, frozen and sublimed to
obtain a lyophilized molecular dispersion.
[0055] For spray dried solid dispersions, the solid dispersion can
be made by a) mixing the compound and polymer in a solvent to
provide a feed solution; and b) spray drying the feed solution to
provide the solid dispersion.
[0056] Spray dried solid dispersions of ledipasvir provide improved
in vivo and in vitro performance and manufacturability/scalability
relative to the other formulation approaches, such as wet and dry
granulation formulations. Ledipasvir can be provided either as the
free base, D-tartrate salt, crystalline acetone solvate, or other
solvate as described herein.
[0057] The selection of the polymer for the solid dispersion is
based on the stability and physical characteristics of the
ledipasvir in the solution. Hypromellose and copovidone solid
dispersions both showed adequate stability and physical
characteristics. Accordingly, in one embodiment, the polymer used
in the solid dispersion is selected from hypromellose and
copovidone. Furthermore, the copovidone-based dispersion increased
in bioavailability more than the equivalent hypromellose-based
formulation (F=30% and 22%, respectively) when prepared at 2:1
API:polymer ratio. Bioavailability of the copovidone-based
formulation was further enhanced by increasing the fraction of
polymer to a 1:1 ratio, resulting in a bioavailability of 35% in
famotidine pretreated dogs.
[0058] In one embodiment, the polymer used in the solid dispersion
of ledipasvir is hydrophilic. Non-limiting examples of hydrophilic
polymers include polysaccharides, polypeptides, cellulose
derivatives such as methyl cellulose, sodium
carboxymethylcellulose, hydroxyethylcellulose, ethylcellulose,
hydroxypropyl methylcellulose acetate-succinate, hydroxypropyl
methylcellulose phthalate, cellulose acetate phthalate,
hydroxypropylcellulose, povidone, copovidone, hypromellose,
pyroxylin, polyethylene oxide, polyvinyl alcohol, and methacrylic
acid copolymers.
[0059] In a further embodiment, the polymer is non-ionic. Non-ionic
polymers showed benefits in screening solubility experiments.
Non-limiting examples of non-ionic polymers include hypromellose,
copovidone, povidone, methyl cellulose, hydroxyethyl cellulose,
hydroxypropyl cellulose, ethylcellulose, pyroxylin, polyethylene
oxide, polyvinyl alcohol, polyethylene glycol, and polyvinyl
caprolactam-polyvinyl acetate-polyethylene glycol.
[0060] In another embodiment, the polymer is ionic. Examples of
ionic polymers include hydroxypropyl methylcellulose
acetate-succinate, hydroxypropyl methylcellulose phthalate,
cellulose acetate phthalate, and methacrylic acid copolymers.
[0061] In a further embodiment, the polymer is selected from the
group consisting of hypromellose, copovidone, and povidone.
Hypromellose and copovidone solid dispersions both showed adequate
stability and physical characteristics. A copovidone-based
dispersion increased bioavailability more than the equivalent
hypromellose-based formulation (F=30% and 22%, respectively) when
spray dried at 2:1 ledipasvir:polymer ratio (data not shown).
Accordingly, in a specific embodiment, the polymer is
copovidone.
[0062] In certain embodiments, the weight ratio of ledipasvir to
polymer is from about 5:1 to about 1:5. In further embodiments, the
weight ratio of ledipasvir to polymer is about 5:1 to about 1:4, or
from about 5:1 to about 1:3, or from about 5:1 to about 1:2, or
from about 2:1 to about 1:2, or from about 2:1 to about 1:1. In a
specific embodiment, the weight ratio of ledipasvir to polymer is
about 1:1. In another embodiment, the weight ratio of ledipasvir to
polymer is about 2:1. In further embodiments, the weight ratio of
ledipasvir to polymer is about 5:1, 1:4, 1:3, or 1:2. Increasing
the fraction of polymer to a 1:1 ratio may, in some instances,
result in an increased bioavailability. For example, a 1:1 ratio of
ledipasvir:copovidone resulted in increased bioavailability (F=35%)
in famotidine pretreated dogs.
[0063] The solid dispersion comprising ledipasvir may be present in
the pharmaceutical composition in a therapeutically effective
amount. In some embodiments, the pharmaceutical compositions
comprises from about 1% to about 50% w/w of the solid dispersion of
ledipasvir. In further embodiments, the composition comprises from
about 5% to about 40% w/w, or from about 5% to about 35% w/w, or
from about 5% to about 30% w/w, or from about 10% to about 30% w/w,
or from about 10% to about 25% w/w, or from about 15% to about 20%
w/w of the solid dispersion of ledipasvir. In further embodiments,
the pharmaceutical composition comprises about 1% w/w, about 5%
w/w, about 10% w/w, about 20% w/w, about 25% w/w, about 30% w/w,
about 35% w/w, or about 40% w/w of the solid dispersion of
ledipasvir. In a specific embodiment, the pharmaceutical
composition comprises about 18% w/w of the solid dispersion of
ledipasvir.
[0064] Ledipasvir may be present in the pharmaceutical composition
in a therapeutically effective amount. In some embodiments, the
pharmaceutical compositions comprises from about 1% to about 50%
w/w of ledipasvir. In further embodiments, the composition
comprises from about 1% to about 40% w/w, or from about 1% to about
30% w/w, or from about 1% to about 20% w/w, or from about 5% to
about 15% w/w, or from about 7% to about 12% w/w of ledipasvir. In
further embodiments, the pharmaceutical composition comprises about
1% w/w, about 3% w/w, about 5% w/w, about 7% w/w, about 11% w/w,
about 13% w/w, about 15% w/w, about 17% w/w, about 20% w/w, about
23% w/w, about 25% w/w, or about 28% w/w, or about 30% w/w of
ledipasvir. In a specific embodiment, the pharmaceutical
composition comprises about 9% w/w of ledipasvir.
[0065] As noted above, after the ledipasvir is mixed with the
polymer, the mixture can then be solubilized in a solvent. It is
within the skill of those in the art to select an appropriate
solvent based on the drug and/or polymer properties such as
solubility, glass transition temperature, viscosity, and molecular
weight. Acceptable solvents include but are not limited to, water,
acetone, methyl acetate, ethyl acetate, chlorinated solvents,
ethanol, dichloromethane, and methanol. In one embodiment, the
solvent is selected from the group consisting of ethanol,
dichloromethane, and methanol. In a further embodiment, the solvent
is ethanol or methanol. In a specific embodiment, the solvent is
ethanol.
[0066] Upon solubilization of the compound and polymer mixture with
the solvent, the mixture may then be spray dried. Spray drying is a
well known process wherein a liquid feedstock is dispersed into
droplets into a drying chamber along with a heated process gas
stream to aid in solvent removal and to produce a powder product.
Suitable spray drying parameters are known in the art, and it is
within the knowledge of a skilled artisan in the field to select
appropriate parameters for spray drying. The target feed
concentration is generally about 10 to about 50% with a target of
about 20% and a viscosity of about 15 to about 300 cP. The inlet
temperature of the spray dry apparatus is typically about
50-190.degree. C., while the outlet temperature is about
30-90.degree. C. The two fluid nozzle and hydrolic pressure nozzle
can be used to spray dry ledipasvir. The two fluid nozzle gas flow
can be about 1-10 kg/hr, the hydrolic pressure nozzle flow can be
about 15-300 kg/hr, and the chamber gas flow may be about 25-2500
kg/hr. The spray-dried material typically has particle size (D90)
under 80 .mu.m. In some instances, a milling step may be used, if
desired to further reduce the particle size. Further descriptions
of spray drying methods and other techniques for forming amorphous
dispersions are provided in U.S. Pat. No. 6,763,607 and U.S. Pat.
Pub. No. 2006-0189633, the entirety of each of which is
incorporated herein by reference.
[0067] Spray drying out of ethanol resulted in high yields (88, 90,
92, 95, 97, 98, 99%) across a wide range of spray-drying outlet
temperatures (30-90.degree. C.) with no material accumulation on
the spray dry chamber, and the yields obtained from spray drying
out of DCM were 60%, 78%, and 44%. Furthermore, ledipasvir
demonstrated good chemical stability in the ethanolic feed
solution.
B. Sofosbuvir
[0068] Sofosbuvir has previously been described in U.S. Pat. No.
7,964,580 and U.S. Publication Nos: 2010/0016251, 2010/0298257,
2011/0251152 and 2012/0107278. Sofosbuvir is provided as
substantially crystalline in the pharmaceutical compositions
described herein. Examples of preparing crystalline forms of
sofosbuvir are disclosed in U.S. Publication Nos: 2010/0298257 and
2011/0251152, both of which are incorporated by reference.
Crystalline forms, Forms 1-6, of sofosbuvir are described in U.S.
Publication Nos.: 2010/0298257 and 2011/0251152, both of which are
incorporated by reference. Forms 1-6 of sofosbuvir have the
following characteristic X-ray powder diffraction (XRPD) pattern
2.theta.-values measured according to the XRPD methods disclosed
therein: [0069] (1) 2.theta.-reflections (.degree..+-.0.2.theta.)
at about: 7.5, 9.6, and 18.3 (Form 1); [0070] (2)
2.theta.-reflections (.degree..+-.0.2.theta.) at about: 5.0, 7.3,
and 18.1 (Form 1); [0071] (3) 2.theta.-reflections
(.degree..+-.0.2.theta.) at about: 6.9, 24.7, and 25.1 (Form 2);
[0072] (4) 2.theta.-reflections (.degree..+-.0.2.theta.) at about:
19.7, 20.6, and 24.6 (Form 3); [0073] (5) 2.theta.-reflections
(.degree..+-.0.2.theta.) at about: 5.0, 6.8, and 24.9 (Form 4);
[0074] (6) 2.theta.-reflections (.degree..+-.0.2.theta.) at about:
5.2, 6.6, and 19.1 (Form 5); and [0075] (7) 2.theta.-reflections
(.degree..+-.0.2.theta.) at about: 6.1, 20.1, and 20.8 (Form
6).
[0076] Form 6, as described in the patent publications above, may
be referred to as Form 2, such for example, by the Food and Drug
Administration. Forms 1 and 6 are alternatively characterized by
the following characteristic XRPD pattern 2.theta.-values as
measured according to the methods disclosed in U.S. Pat. Pub. Nos.:
2010/0298257 and 2011/0251152: [0077] (1) 2.theta.-reflections
(.degree.) at about: 5.0 and 7.3 (Form 1); and [0078] (2)
2.theta.-reflections (.degree.) at about: 6.1 and 12.7 (Form
6).
[0079] In one embodiment, the crystalline sofosbuvir has XRPD
2.theta.-reflections (.degree..+-.0.2.theta.) at about: [0080] (1)
7.5, 9.6, and 18.3; (Form 1A) [0081] (2) 5.0, 7.3, and 18.1; (Form
1B) [0082] (3) 6.9, 24.7, and 25.1; (Form 2) [0083] (4) 19.7, 20.6,
and 24.6; (Form 3) [0084] (5) 5.0, 6.8, and 24.9; (Form 4) [0085]
(6) 5.2, 6.6, and 19.1; (Form 5) or [0086] (7) 6.1, 20.1, and 20.8;
(Form 6).
[0087] In certain embodiments, the crystalline sofosbuvir has XRPD
2.theta.-reflections (.degree..+-.0.2.theta.) at about: [0088] (1)
5.2, 7.5, 9.6, 16.7, 18.3, and 22.2 (Form 1); [0089] (2) 5.0, 7.3,
9.4, and 18.1 (Form 1); [0090] (3) 4.9, 6.9, 9.8, 19.8, 20.6, 24.7,
25.1, and 26.1 (Form 2); [0091] (4) 6.9, 9.8, 19.7, 20.6, and 24.6
(Form 3); [0092] (5) 5.0, 6.8, 19.9, 20.6, 20.9, and 24.9 (Form 4);
[0093] (6) 5.2, 6.6, 7.1, 15.7, 19.1, and 25.0 (Form 5); or [0094]
(7) 6.1, 8.2, 10.4, 12.7, 17.2, 17.7, 18.0, 18.8, 19.4, 19.8, 20.1,
20.8, 21.8, and 23.3 (Form 6).
[0095] In a further embodiment, crystalline sofosbuvir has XRPD
2.theta.-reflections (.degree..+-.0.2.theta.) at about: 6.1, 8.2,
10.4, 12.7, 17.2, 17.7, 18.0, 18.8, 19.4, 19.8, 20.1, 20.8, 21.8,
and 23.3. In yet a further embodiment, crystalline sofosbuvir has
XRPD 2.theta.-reflections (.degree..+-.0.2.theta.) at about: 6.1
and 12.7.
[0096] Sofosbuvir may be present in the pharmaceutical composition
in a therapeutically effective amount. In some embodiments, the
pharmaceutical compositions comprises from about 10% to about 70%
w/w of sofosbuvir. In further embodiments, the composition
comprises from about 15% to about 65% w/w, or from about 20% to
about 60% w/w, or from about 25% to about 55% w/w, or from about
30% to about 50% w/w, or from about 35% to about 45% w/w of
sofosbuvir. In further embodiments, the pharmaceutical composition
comprises about 10% w/w, about 15% w/w, about 20% w/w, about 25%
w/w, about 30% w/w, about 35% w/w, about 45% w/w, about 50% w/w,
about 55% w/w, about 60% w/w, about 65% w/w, or about 70% w/w, or
about 75% w/w. In a specific embodiment, the pharmaceutical
composition comprises about 40% w/w of sofosbuvir.
C. Excipients
[0097] The pharmaceutical compositions provided in accordance with
the present disclosure are usually administered orally. This
disclosure therefore provides pharmaceutical compositions that
comprise a solid dispersion comprising ledipasvir as described
herein and one or more pharmaceutically acceptable excipients or
carriers including but not limited to, inert solid diluents and
fillers, diluents, including sterile aqueous solution and various
organic solvents, permeation enhancers, solubilizers,
disintegrants, lubricants, binders, glidants, adjuvants, and
combinations thereof. Such compositions are prepared in a manner
well known in the pharmaceutical art (see, e.g., Remington's
Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa.
17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd
Ed. (G. S. Banker & C. T. Rhodes, Eds.).
[0098] The pharmaceutical compositions may be administered in
either single or multiple doses by oral administration.
Administration may be via capsule, tablet, or the like. In one
embodiment, the ledipasvir is in the form of a tablet. In a further
embodiment, the tablet is a compressed tablet. In making the
pharmaceutical compositions that include the solid described
herein, the active ingredient is usually diluted by an excipient
and/or enclosed within such a carrier that can be in the form of a
capsule, tablet, sachet, paper or other container. When the
excipient serves as a diluent, it can be in the form of a solid,
semi-solid or liquid material (as above), which acts as a vehicle,
carrier or medium for the active ingredient.
[0099] The pharmaceutical composition may be formulated for
immediate release or sustained release. A "sustained release
formulation" is a formulation which is designed to slowly release a
therapeutic agent in the body over an extended period of time,
whereas an "immediate release formulation" is an formulation which
is designed to quickly release a therapeutic agent in the body over
a shortened period of time. In some cases the immediate release
formulation may be coated such that the therapeutic agent is only
released once it reached the desired target in the body (e.g. the
stomach). In a specific embodiment, the pharmaceutical composition
is formulated for immediate release.
[0100] The pharmaceutical composition may further comprise
pharmaceutical excipients such as diluents, binders, fillers,
glidants, disintegrants, lubricants, solubilizers, and combinations
thereof. Some examples of suitable excipients are described herein.
When the pharmaceutical composition is formulated into a tablet,
the tablet may be uncoated or may be coated by known techniques
including microencapsulation to delay disintegration and adsorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate alone or with
a wax may be employed.
[0101] In one embodiment, the pharmaceutical composition comprises
a diluent selected from the group consisting of dicalcium
phosphate, cellulose, compressible sugars, dibasic calcium
phosphate dehydrate, lactose, lactose monohydrate, mannitol,
microcrystalline cellulose, starch, tribasic calcium phosphate, and
combinations thereof.
[0102] In further embodiments, the pharmaceutical composition
comprises lactose monohydrate in an amount from about 1 to about
50% w/w, or from about 1 to about 45% w/w, or from about 5 to about
40% w/w, or from about 5 to about 35% w/w, or from about 5 to about
25% w/w, or from about 10 to about 20% w/w. In specific
embodiments, the lactose monohydrate is present at about 5% w/w, at
about 10% w/w, at about 15% w/w, at about 20% w/w, at about 25%
w/w, at about 30% w/w, at about 35% w/w, at about 40% w/w, at about
45% w/w, or at about 50% w/w. In a further specific embodiment, the
lactose monohydrate is in an amount of about 16.5% w/w.
[0103] In yet further embodiments, the pharmaceutical composition
comprises microcrystalline cellulose in an amount from about 1 to
about 40% w/w, or from about 1 to about 35% w/w, or from about 1%
to about 25% w/w, or from about 5 to about 25% w/w, or from about
10 to about 25% w/w, or from about 15 to about 20% w/w. In specific
embodiments, the microcrystalline cellulose is present in an amount
of about 5%, or about 10%, or about 15%, or about 20%, or about
25%, or about 30%, or about 35%, or about 40% w/w. In a further
specific embodiment, the microcrystalline cellulose is in an amount
of about 18% w/w.
[0104] In other embodiments, the pharmaceutical composition
comprises a disintegrant selected from the group consisting of
croscarmellose sodium, crospovidone, microcrystalline cellulose,
modified corn starch, povidone, pregelatinized starch, sodium
starch glycolate, and combinations thereof.
[0105] In certain embodiments, the pharmaceutical composition
comprises croscarmellose sodium in an amount from about 1 to about
20% w/w, or from about 1 to about 15% w/w, or from about 1 to about
10% w/w, or from about 1 to about 8% w/w, or from about 2 to about
8% w/w. In specific embodiments, the croscarmellose sodium is
present in an amount of about 1%, or about 3%, or about 6%, or
about 8%, or about 10%, or about 13%, or about 15% w/w. In a
further specific embodiment, the croscarmellose sodium is in an
amount of about 5% w/w.
[0106] In other embodiments, the pharmaceutical composition
comprises a glidant selected from the group consisting of colloidal
silicon dioxide, talc, starch, starch derivatives, and combinations
thereof.
[0107] In further embodiments, the pharmaceutical composition
comprises colloidal silicon dioxide in an amount from about 0.1 to
about 5% w/w, or from about 0.1 to about 4.5% w/w, or from about
0.1 to about 4% w/w, or from about 0.5 to about 5.0% w/w, or from
about 0.5 to about 3% w/w, or from about 0.5 to about 2% w/w, or
from about 0.5 to about 1.5% w/w. In specific embodiments, the
colloidal silicon dioxide is present in an amount of about 0.1%
w/w, 0.5% w/w, 0.75% w/w, 1.25% w/w, 1.5% w/w, or 2% w/w. In a
further specific embodiment, the colloidal silicon dioxide is
present in an amount of about 1% w/w.
[0108] In other embodiments, the pharmaceutical composition
comprises a lubricant selected from the group consisting of calcium
stearate, magnesium stearate, polyethylene glycol, sodium stearyl
fumarate, stearic acid, talc, and combinations thereof.
[0109] In further embodiments, the pharmaceutical composition
comprises magnesium stearate in an amount from about 0.1 to about
3% w/w, or from about 0.1 to about 2.5% w/w, or from about 0.5 to
about 3% w/w, or from about 0.5 to about 2.5% w/w, or from about
0.5 to about 2% w/w, or from about 1 to about 3% w/w, or from about
1 to about 2% w/w. In specific embodiments, the magnesium stearate
is present in an amount of about 0.1%, or about 0.5, or about 1%,
or about 2%, or about 2.5%, or about 3% w/w. In a further specific
embodiment, the magnesium stearate is in an amount of about 1.5%
w/w.
[0110] In one embodiment, the pharmaceutical composition comprises
a) about 30 to about 50% w/w of sofosbuvir and b) about 5 to about
35% w/w of the solid dispersion comprising ledipasvir. In a related
embodiment, the composition comprises a) about 40% w/w of
sofosbuvir and b) about 18% w/w of the solid dispersion comprising
ledipasvir. In yet a further related embodiment, the composition
further comprises a) about 5 to about 25% w/w lactose monohydrate,
b) about 5 to about 25% w/w microcrystalline cellulose, c) about 1
to about 10% w/w croscarmellose sodium, d) about 0.5 to about 3%
w/w colloidal silicon dioxide, and e) about 0.1 to about 3% w/w
magnesium stearate. In a further embodiment, the pharmaceutical
composition comprises a) about 40% w/w of sofosbuvir, b) about 18%
w/w of the solid dispersion comprising ledipasvir, c) about 16.5%
w/w lactose monohydrate, d) about 18% w/w microcrystalline
cellulose, e) about 5% w/w croscarmellose sodium, f) about 1% w/w
colloidal silicon dioxide, and g) about 1.5% w/w magnesium
stearate.
3. Pharmaceutical Dosage Forms
[0111] The disclosure provides for tablets, pills, and the like,
comprising the pharmaceutical compositions or dosage forms
described herein. The tablets or pills of the present disclosure
may be coated to provide a dosage form affording the advantage of
prolonged action or to protect from the acid conditions of the
stomach. The tablets may also be formulated for immediate release
as previously described. In certain embodiments, the tablet
comprises a film coating. A film coating is useful for limiting
photolytic degradation. Suitable film coatings are selected by
routine screening of commercially available preparations. In one
embodiment, the film coating is a polyvinylalcohol-based
coating.
[0112] The tablets may be formulated into a monolayer or bilayer
tablet. Typically, monolayer tablet comprise the active ingredients
(i.e., ledipasvir and sofosbuvir) co-mixed in a single uniform
layer. For making monolayer tablets, exemplary methods include, but
are not limited to coblend (or bi-granulation) and codry
granulation. Coblend granulation is a multi-step process consisting
of separate dry granulations for each active ingredient with
excipients followed by the blending of the two granulations
together. Codry granulation consisted of dry granulating both
active ingredients and excipients together.
[0113] Bilayer tablets comprise the active ingredients (i.e.,
ledipasvir and sofosbuvir) in separate layers and can be made by
making a blend comprising excipients and one active ingredient
(i.e., ledipasvir), and making a separate blend comprising the
second active ingredient (i.e., sofosbuvir) and excipients. One
blend may then be precompressed, and the second blend may then be
added on top of the first precompressed blends. The resulting
tablet comprises two separate layers, each layer comprising a
different active ingredient.
[0114] In one embodiment, the tablet comprises a) about 30 to about
50% w/w of sofosbuvir and b) about 10 to about 40% w/w of the solid
dispersion comprising ledipasvir. In a related embodiment, the
tablet comprises a) about 40% w/w of sofosbuvir and b) about 18%
w/w of the solid dispersion comprising ledipasvir. In a further
embodiment, the tablet comprises a) about 300 to about 500 mg of
sofosbuvir and b) about 50 to about 130 mg of ledipasvir. In a yet
further embodiment, the tablet comprises a) about 400 mg of
sofosbuvir and b) about 90 mg of ledipasvir. In related embodiment,
the tablet further comprises a) about 5 to about 25% w/w lactose
monohydrate, b) about 5 to about 25% w/w microcrystalline
cellulose, c) about 1 to about 10% w/w croscarmellose sodium, d)
about 0.5 to about 3% w/w colloidal silicon dioxide, and e) about
0.1 to about 3% w/w magnesium stearate.
[0115] In some embodiments, the pharmaceutical compositions as
described herein are formulated in a unit dosage or pharmaceutical
dosage form. The term "unit dosage forms" or "pharmaceutical dosage
forms" refers to physically discrete units suitable as unitary
dosages for human patients and other mammals, each unit containing
a predetermined quantity of active material calculated to produce
the desired therapeutic effect, in association with a suitable
pharmaceutical excipient (e.g., a tablet or capsule). The compounds
are generally administered in a pharmaceutically effective amount.
In some embodiments, each dosage unit contains from 3 mg to 2 g of
ledipasvir. In other embodiments, the pharmaceutical dosage form
comprises from about 3 to about 360 mg, or about 10 to about 200
mg, or about 10 to about 50 mg, or about 20 to about 40 mg, or
about 25 to about 35 mg, or about 40 to about 140 mg, or about 50
to about 130 mg, or about 60 to about 120 mg, or about 70 to about
110 mg, or about 80 to about 100 mg. In specific embodiments, the
pharmaceutical dosage form comprises about 40, or about 45, or
about 50, or about 55, or about 60, or about 70, or about 80, or
about 100, or about 120, or about 140, or about 160, or about 180,
or about 200, or about 220 mg of ledipasvir. In a further specific
embodiment, the pharmaceutical dosage form comprises about 90 mg of
ledipasvir. In yet a further specific embodiment, the
pharmaceutical dosage form comprises about 30 mg of ledipasvir.
[0116] In other embodiments, the pharmaceutical dosage form
comprises from about 1 mg to about 3 g of sofosbuvir. In other
embodiments, the pharmaceutical dosage form comprises from about 1
to about 800 mg, or about 100 to about 700 mg, or about 200 to
about 600 mg, or about 300 to about 500 mg, or about 350 to about
450 mg, of sofosbuvir. In specific embodiments, the pharmaceutical
dosage form comprises about 50, or about 100, or about 150, or
about 200, or about 250, or about 300, or about 350, or about 450,
or about 500, or about 550, or about 600, or about 650, or about
700, or about 750, or about 800 mg of sofosbuvir. In a further
specific embodiment, the pharmaceutical dosage form comprises about
400 mg of sofosbuvir. It will be understood, however, that the
amount of ledipasvir and/or sofosbuvir actually administered
usually will be determined by a physician, in the light of the
relevant circumstances, including the condition to be treated, the
chosen route of administration, the actual compound administered
and its relative activity, the age, weight and response of the
individual patient, the severity of the patient's symptoms, and the
like.
[0117] In a specific embodiment, the pharmaceutical dosage form
comprises about 400 mg of sofosbuvir and about 90 mg of
ledipasvir.
[0118] In one embodiment, the pharmaceutical composition, or
alternatively, the pharmaceutical dosage form or tablet comprises
about 90 mg of amorphous ledipasvir formulated in a solid
dispersion comprising a polymer:ledipasvir ratio of 1:1, about 400
mg crystalline sofosbuvir, lactose monohydrate in an amount from
about 5 to about 25% w/w, microcrystalline cellulose in an amount
from about 5 to about 25% w/w, croscarmellose sodium in an amount
from about 1 to about 10% w/w, colloidal silicon dioxoide in an
amount from about 0.5 to about 3% w/w, and magnesium stearate in an
amount from about 0.1 to about 3% w/w. In one embodiment, the
polymer is copovidone.
[0119] In further embodiments, the pharmaceutical composition,
pharmaceutical dosage form, or tablet as described herein is free
of negative drug-drug interactions. In a related embodiment, the
pharmaceutical composition, pharmaceutical dosage form, or tablet
is free of negative drug-drug interactions with acid suppressive
therapies. In a further embodiment, the pharmaceutical composition,
pharmaceutical dosage form, or tablet as described herein is
administrable without regard to food and with or without regard to
the patient being on an acid-suppressive therapy.
4. Methods of Use
[0120] The solid dispersions, pharmaceutical compositions,
pharmaceutical dosage forms, and tablets of ledipasvir and
sofosbuvir as described herein are administered to a patient
suffering from hepatitis C virus (HCV) in a daily dose by oral
administration. In one embodiment, the patient is human.
[0121] Previously, ledipasvir had been demonstrated to have a
negative food effect when administered alone. Unexpectedly, the
combination treatment of ledipasvir and sofosbuvir does not exhibit
a negative food effect. Accordingly, the administration of the
pharmaceutical composition comprising sofosbuvir and ledipasvir can
be taken without regard to food.
[0122] In some embodiments, the combination composition achieved a
reduced food effect. In some aspects, the composition achieves a
first exposure, when administered to a patient following a meal,
that is no more than 25%, or alternatively not more than 20%, 15%
or 10%, lower than a second exposure when administered to the
patient not following a meal. The exposures can be measured as
C.sub.max, AUC.sub.last or AUC.sub.inf. In some aspects, the
administration is carried out within four, three, two or one hours
following the meal.
[0123] In one embodiment, the solid dispersions, pharmaceutical
compositions, pharmaceutical dosage forms, and tablets of
ledipasvir and sofosbuvir as described herein are effective in
treating one or more of genotype 1 HCV infected patients, genotype
2 HCV infected patients, genotype 3 HCV infected patients, genotype
4 HCV infected patients, genotype 5 HCV infected patients, and/or
genotype 6 HCV infected patients. In one embodiment, the solid
dispersions, pharmaceutical compositions, pharmaceutical dosage
forms, and tablets of ledipasvir and sofosbuvir as described herein
are effective in treating genotype 1 HCV infected patients,
including genotype 1a and/or genotype 1b. In another embodiment,
the solid dispersions, pharmaceutical compositions, pharmaceutical
dosage forms, and tablets of ledipasvir and sofosbuvir as described
herein are effective in treating genotype 2 HCV infected patients,
including genotype 2a, genotype 2b, genotype 2c and/or genotype 2d.
In another embodiment, the solid dispersions, pharmaceutical
compositions, pharmaceutical dosage forms, and tablets of
ledipasvir and sofosbuvir as described herein are effective in
treating genotype 3 HCV infected patients, including genotype 3a,
genotype 3b, genotype 3c, genotype 3d, genotype 3e and/or genotype
3f. In another embodiment, the solid dispersions, pharmaceutical
compositions, pharmaceutical dosage forms, and tablets of
ledipasvir and sofosbuvir as described herein are effective in
treating genotype 4 HCV infected patients, including genotype 4a,
genotype 4b, genotype 4c, genotype 4d, genotype 4e, genotype 4f,
genotype 4g, genotype 4h, genotype 4i and/or genotype 4j. In
another embodiment, the solid dispersions, pharmaceutical
compositions, pharmaceutical dosage forms, and tablets of
ledipasvir and sofosbuvir as described herein are effective in
treating genotype 5 HCV infected patients, including genotype 5a.
In another embodiment, the solid dispersions, pharmaceutical
compositions, pharmaceutical dosage forms, and tablets of
ledipasvir and sofosbuvir as described herein are effective in
treating genotype 6 HCV infected patients, including genotype 6a.
In one embodiment, the compositions are pangenotypic, meaning they
are useful across all genotypes and drug resistant mutants
thereof.
[0124] In some embodiments, the pharmaceutical composition,
pharmaceutical dosage form, or tablet of ledipasvir and sofosbuvir
as described herein is administered, either alone or in combination
with one or more therapeutic agent(s) for treating HCV (such as a
HCV NS3 protease inhibitor or an inhibitor of HCV NS5B polymerase),
for about 24 weeks, for about 16 weeks, or for about 12 weeks, or
less. In further embodiments, the pharmaceutical composition,
pharmaceutical dosage form, or tablet of ledipasvir and sofosbuvir
is administered, either alone or in combination with one or more
therapeutic agent(s) for treating HCV (such as a HCV NS3 protease
inhibitor or an inhibitor of HCV NS5B polymerase), for about 24
weeks or less, about 22 weeks or less, about 20 weeks or less,
about 18 weeks or less, about 16 weeks or less, about 12 weeks or
less, about 10 weeks or less, about 8 weeks or less, or about 6
weeks or less or about 4 weeks or less. The pharmaceutical
composition, pharmaceutical dosage form, or tablet may be
administered once daily, twice daily, once every other day, two
times a week, three times a week, four times a week, or five times
a week.
[0125] In further embodiments, a sustained virologic response is
achieved at about 4 weeks, 6 weeks, 8 weeks, 12 weeks, or 16 weeks,
or at about 20 weeks, or at about 24 weeks, or at about 4 months,
or at about 5 months, or at about 6 months, or at about 1 year, or
at about 2 years.
[0126] In one embodiment, the daily dose is 90 mg of ledipasvir and
400 mg of sofosbuvir administered in the form of a tablet. In a
further embodiment, the daily dose is a tablet comprising a) about
30 to about 50% w/w of sofosbuvir, b) about 10 to about 40% w/w of
the solid dispersion comprising ledipasvir, c) about 5 to about 25%
w/w lactose monohydrate, d) about 5 to about 25% w/w
microcrystalline cellulose, e) about 1 to about 10% w/w
croscarmellose sodium, f) about 0.5 to about 3% w/w colloidal
silicon dioxide, and g) about 0.1 to about 3% w/w magnesium
stearate.
[0127] In further embodiments, the patient is also suffering from
cirrhosis. In yet a further embodiment, the patient is not
suffereing from cirrhosis.
5. Combination Therapy
[0128] In the methods described herein, the method can further
comprise the administration of another therapeutic agent for
treating HCV and other conditions such as HIV infections. In one
embodiment, non-limiting examples of suitable additional
therapeutic agents include one or more interferons, ribavirin or
its analogs, HCV NS3 protease inhibitors, alpha-glucosidase 1
inhibitors, hepatoprotectants, nucleoside or nucleotide inhibitors
of HCV NS5B polymerase, non-nucleoside inhibitors of HCV NS5B
polymerase, HCV NS5A inhibitors, TLR-7 agonists, cyclophillin
inhibitors, HCV IRES inhibitors, pharmacokinetic enhancers, and
other drugs or therapeutic agents for treating HCV.
[0129] More specifically, the additional therapeutic agent may be
selected from the group consisting of:
[0130] 1) interferons, e.g., pegylated rIFN-alpha 2b (PEG-Intron),
pegylated rIFN-alpha 2a (Pegasys), rIFN-alpha 2b (Intron A),
rIFN-alpha 2a (Roferon-A), interferon alpha (MOR-22, OPC-18,
Alfaferone, Alfanative, Multiferon, subalin), interferon alfacon-1
(Infergen), interferon alpha-nl (Wellferon), interferon alpha-n3
(Alferon), interferon-beta (Avonex, DL-8234), interferon-omega
(omega DUROS, Biomed 510), albinterferon alpha-2b (Albuferon), IFN
alpha-2b XL, BLX-883 (Locteron), DA-3021, glycosylated interferon
alpha-2b (AVI-005), PEG-Infergen, PEGylated interferon lambda-1
(PEGylated IL-29), and belerofon;
[0131] 2) ribavirin and its analogs, e.g., ribavirin (Rebetol,
Copegus), and taribavirin (Viramidine);
[0132] 3) HCV NS3 protease inhibitors, e.g., boceprevir
(SCH-503034, SCH-7), telaprevir (VX-950), TMC435350, BI-1335,
BI-1230, MK-7009, VBY-376, VX-500, GS-9256, GS-9451, BMS-605339,
PHX-1766, AS-101, YH-5258, YH5530, YH5531, ABT-450, ACH-1625,
ITMN-191, MK5172, MK6325, and MK2748;
[0133] 4) alpha-glucosidase 1 inhibitors, e.g., celgosivir
(MX-3253), Miglitol, and UT-231B;
[0134] 5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738,
GS-9450 (LB-84451), silibilin, and MitoQ;
[0135] 6) nucleoside or nucleotide inhibitors of HCV NS5B
polymerase, e.g., R1626, R7128 (R4048), IDX184, IDX-102, BCX-4678,
valopicitabine (NM-283), MK-0608, and INX-189 (now BMS986094);
[0136] 7) non-nucleoside inhibitors of HCV NS5B polymerase, e.g.,
PF-868554, VCH-759, VCH-916, JTK-652, MK-3281, GS-9190, VBY-708,
VCH-222, A848837, ANA-598, GL60667, GL59728, A-63890, A-48773,
A-48547, BC-2329, VCH-796 (nesbuvir), GSK625433, BILN-1941,
XTL-2125, ABT-072, ABT-333, GS-9669, PSI-7792, and GS-9190;
[0137] 8) HCV NS5A inhibitors, e.g., AZD-2836 (A-831), BMS-790052,
ACH-3102, ACH-2928, MK8325, MK4882, MK8742, PSI-461, IDX719,
ABT-267, and A-689;
[0138] 9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773,
ANA-975, AZD-8848 (DSP-3025), and SM-360320;
[0139] 10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and
NIM811;
[0140] 11) HCV IRES inhibitors, e.g., MCI-067;
[0141] 12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452,
PF-4194477, TMC-41629, GS-9350, GS-9585, and roxythromycin; and
[0142] 13) other drugs for treating HCV, e.g., thymosin alpha 1
(Zadaxin), nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17
(altirex), KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000,
civacir, GI-5005, XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA
971, NOV-205, tarvacin, EHC-18, VGX-410C, EMZ-702, AVI 4065,
BMS-650032, BMS-791325, Bavituximab, MDX-1106 (ONO-4538),
Oglufanide, and VX-497 (merimepodib).
[0143] More specifically, the additional therapeutic agent may be
combined with one or more compounds selected from the group
consisting of non-nucleoside inhibitors of HCV NS5B polymerase
(ABT-072 and ABT-333), HCV NS5A inhibitors (ACH-3102 and ACH-2928)
and HCV NS3 protease inhibitors (ABT-450 and ACH-125).
[0144] In another embodiment, the therapeutic agent used in
combination with the pharmaceutical compositions as described
herein can be any agent having a therapeutic effect when used in
combination with the pharmaceutical compositions as described
herein. For example, the therapeutic agent used in combination with
the pharmaceutical compositions as described herein can be
interferons, ribavirin analogs, NS3 protease inhibitors, NS5B
polymerase inhibitors, alpha-glucosidase 1 inhibitors,
hepatoprotectants, non-nucleoside inhibitors of HCV, and other
drugs for treating HCV.
[0145] In another embodiment, the additional therapeutic agent used
in combination with the pharmaceutical compositions as described
herein is a cyclophillin inhibitor, including for example, a
cyclophilin inhibitor disclosed in WO2013/185093. Non-limiting
examples include one or more compounds selected from the group
consisiting of:
##STR00003## ##STR00004##
and stereoisomers and mixtures of stereoisomers thereof.
[0146] In another embodiment, the additional therapeutic agent used
in combination with the pharmaceutical compositions as described
herein is a non-nucleoside inhibitor of HCV NS5B polymerase. A
non-limiting example includes Compound E (as described below).
[0147] Examples of additional anti-HCV agents which can be combined
with the compositions provided herein include, without limitation,
the following:
[0148] A. interferons, for example, pegylated rIFN-alpha 2b
(PEG-Intron), pegylated rIFN-alpha 2a (Pegasys), rIFN-alpha 2b
(Intron A), rIFN-alpha 2a (Roferon-A), interferon alpha (MOR-22,
OPC-18, Alfaferone, Alfanative, Multiferon, subalin), interferon
alfacon-1 (Infergen), interferon alpha-nl (Wellferon), interferon
alpha-n3 (Alferon), interferon-beta (Avonex, DL-8234),
interferon-omega (omega DUROS, Biomed 510), albinterferon alpha-2b
(Albuferon), IFN alpha XL, BLX-883 (Locteron), DA-3021,
glycosylated interferon alpha-2b (AVI-005), PEG-Infergen, PEGylated
interferon lambda (PEGylated IL-29), or belerofon, IFN alpha-2b XL,
rIFN-alpha 2a, consensus IFN alpha, infergen, rebif, pegylated
IFN-beta, oral interferon alpha, feron, reaferon, intermax alpha,
r-IFN-beta, and infergen+actimmuneribavirin and ribavirin analogs,
e.g., rebetol, copegus, VX-497, and viramidine (taribavirin);
[0149] B. NS5A inhibitors, for example, Compound B (described
below), Compound C (described below), ABT-267, Compound D
(described below), JNJ-47910382, daclatasvir (BMS-790052), ABT-267,
MK-8742, EDP-239, IDX-719, PPI-668, GSK-2336805, ACH-3102, A-831,
A-689, AZD-2836 (A-831), AZD-7295 (A-689), and BMS-790052;
[0150] C. NS5B polymerase inhibitors, for example, Compound E
(described below), Compound F (described below), ABT-333, Compound
G (described below), ABT-072, Compound H (described below),
tegobuvir (GS-9190), GS-9669, TMC647055, setrobuvir (ANA-598),
filibuvir (PF-868554), VX-222, IDX-375, IDX-184, IDX-102,
BI-207127, valopicitabine (NM-283), PSI-6130 (R1656), PSI-7851,
BCX-4678, nesbuvir (HCV-796), BILB 1941, MK-0608, NM-107, R7128,
VCH-759, GSK625433, XTL-2125, VCH-916, JTK-652, MK-3281, VBY-708,
A848837, GL59728, A-63890, A-48773, A-48547, BC-2329, BMS-791325,
and BILB-1941;
[0151] D. NS3 protease inhibitors, for example, Compound I,
Compound J, Compound K, ABT-450, Compound L (described below),
simeprevir (TMC-435), boceprevir (SCH-503034), narlaprevir
(SCH-900518), vaniprevir (MK-7009), MK-5172, danoprevir (ITMN-191),
sovaprevir (ACH-1625), neceprevir (ACH-2684), Telaprevir (VX-950),
VX-813, VX-500, faldaprevir (BI-201335), asunaprevir (BMS-650032),
BMS-605339, VBY-376, PHX-1766, YH5531, BILN-2065, and
BILN-2061;
[0152] E. alpha-glucosidase 1 inhibitors, for example, celgosivir
(MX-3253), Miglitol, and UT-231B;
[0153] F hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and
LB-84451;
[0154] G. non-nucleoside inhibitors of HCV, e.g., benzimidazole
derivatives, benzo-1,2,4-thiadiazine derivatives, and phenylalanine
derivatives; and
[0155] H. other anti-HCV agents, e.g., zadaxin, nitazoxanide
(alinea), BIVN-401 (virostat), DEBIO-025, VGX-410C, EMZ-702, AVI
4065, bavituximab, oglufanide, PYN-17, KPE02003002, actilon
(CPG-10101), KRN-7000, civacir, GI-5005, ANA-975, XTL-6865, ANA
971, NOV-205, tarvacin, EHC-18, and NIM811.
[0156] Compound B is an NS5A inhibitor and is represented by the
following chemical structure:
##STR00005##
[0157] Compound C is an NS5A inhibitor and is represented by the
following chemical structure:
##STR00006##
[0158] Compound D is an NS5A inhibitor and is represented by the
following chemical structure:
##STR00007##
[0159] See U.S. Publication No. 2013/0102525 and references cited
therein.
[0160] Compound E is an NS5B Thumb II polymerase inhibitor and is
represented by the following chemical structure:
##STR00008##
[0161] Compound F is a nucleotide inhibitor prodrug designed to
inhibit replication of viral RNA by the HCV NS5B polymerase, and is
represented by the following chemical structure:
##STR00009##
[0162] Compound G is an HCV polymerase inhibitor and is represented
by the following structure:
##STR00010##
[0163] See U.S. Publication No. 2013/0102525 and references
therein.
[0164] Compound H is an HCV polymerase inhibitor and is represented
by the following structure:
##STR00011##
[0165] See U.S. Publication No. 2013/0102525 and references
therein.
[0166] Compound I is an HCV protease inhibitor and is represented
by the following chemical structure:
##STR00012##
[0167] See U.S. Publication No. 2014/0017198 and references
therein.
[0168] Compound J is an HCV protease inhibitor and is represented
by the following chemical structure:
##STR00013##
[0169] See U.S. Pat. No. 8,178,491 and references therein.
[0170] Compound K is an HCV protease inhibitor and is represented
by the following chemical structure:
##STR00014##
[0171] Compound L is an HCV protease inhibitor and is represented
by the following chemical structure:
##STR00015##
[0172] See U.S. Publication No. 2013/0102525 and references
therein.
[0173] In one embodiment, the additional therapeutic agent used in
combination with the pharmaceutical compositions as described
herein is a HCV NS3 protease inhibitor. Non-limiting examples
include one or more compounds selected from the group consisiting
of:
##STR00016##
[0174] In another embodiment, the present application is provided a
method of treating hepatitis C in a human patient in need thereof
comprising administering to the patient a therapeutically effective
amount of a pharmaceutical composition as described herein and an
additional therapeutic selected from the group consisting of
pegylated rIFN-alpha 2b, pegylated rIFN-alpha 2a, rIFN-alpha 2b,
IFN alpha-2b XL, rIFN-alpha 2a, consensus IFN alpha, infergen,
rebif, locteron, AVI-005, PEG-infergen, pegylated IFN-beta, oral
interferon alpha, feron, reaferon, intermax alpha, r-IFN-beta,
infergen+actimmune, IFN-omega with DUROS, albuferon, rebetol,
copegus, levovirin, VX-497, viramidine (taribavirin), A-831, A-689,
NM-283, valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB
1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433,
XTL-2125, SCH-503034 (SCH-7), VX-950 (Telaprevir), ITMN-191, and
BILN-2065, MX-3253 (celgosivir), UT-231B, IDN-6556, ME 3738, MitoQ,
and LB-84451, benzimidazole derivatives, benzo-1,2,4-thiadiazine
derivatives, and phenylalanine derivatives, zadaxin, nitazoxanide
(alinea), BIVN-401 (virostat), DEBIO-025, VGX-410C, EMZ-702, AVI
4065, bavituximab, oglufanide, PYN-17, KPE02003002, actilon
(CPG-10101), KRN-7000, civacir, GI-5005, ANA-975 (isatoribine),
XTL-6865, ANA 971, NOV-205, tarvacin, EHC-18, and NIM811 and a
pharmaceutically acceptable carrier or excipient.
[0175] In yet another embodiment, the present application provides
a combination pharmaceutical agent comprising:
[0176] a) a first pharmaceutical composition comprising an
effective amount of wherein ledipasvir is substantially amorphous;
and an effective amount of sofosbuvir wherein sofosbuvir is
substantially crystalline as described herein and
[0177] b) a second pharmaceutical composition comprising at least
one additional therapeutic agent selected from the group consisting
of HIV protease inhibiting compounds, HIV non-nucleoside inhibitors
of reverse transcriptase, HIV nucleoside inhibitors of reverse
transcriptase, HIV nucleotide inhibitors of reverse transcriptase,
HIV integrase inhibitors, gp41 inhibitors, CXCR4 inhibitors, gp120
inhibitors, CCR5 inhibitors, interferons, ribavirin analogs, NS3
protease inhibitors, alpha-glucosidase 1 inhibitors,
hepatoprotectants, non-nucleoside inhibitors of HCV, and other
drugs for treating HCV, and combinations thereof.
[0178] The additional therapeutic agent may be one that treats
other conditions such as HIV infections. Accordingly, the
additional therapeutic agent may be a compound useful in treating
HIV, for example HIV protease inhibiting compounds, non-nucleoside
inhibitors of HIV reverse transcriptase, HIV nucleoside inhibitors
of reverse transcriptase, HIV nucleotide inhibitors of reverse
transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4
inhibitors, gp120 inhibitors, CCR5 inhibitors, interferons,
ribavirin analogs, NS3 protease inhibitors, NS5b polymerase
inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants,
non-nucleoside inhibitors of HCV, and other drugs for treating
HCV.
[0179] More specifically, the additional therapeutic agent may be
selected from the group consisting of
[0180] 1) HIV protease inhibitors, e.g., amprenavir, atazanavir,
fosamprenavir, indinavir, lopinavir, ritonavir,
lopinavir+ritonavir, nelfinavir, saquinavir, tipranavir,
brecanavir, darunavir, TMC-126, TMC-114, mozenavir (DMP-450),
JE-2147 (AG1776), AG1859, DG35, L-756423, R00334649, KNI-272,
DPC-681, DPC-684, and GW640385X, DG17, PPL-100,
[0181] 2) a HIV non-nucleoside inhibitor of reverse transcriptase,
e.g., capravirine, emivirine, delaviridine, efavirenz, nevirapine,
(+) calanolide A, etravirine, GW5634, DPC-083, DPC-961, DPC-963,
MIV-150, and TMC-120, TMC-278 (rilpivirine), efavirenz, BILR 355
BS, VRX 840773, UK-453,061, RDEA806,
[0182] 3) a HIV nucleoside inhibitor of reverse transcriptase,
e.g., zidovudine, emtricitabine, didanosine, stavudine,
zalcitabine, lamivudine, abacavir, amdoxovir, elvucitabine,
alovudine, MIV-210, racivir (.+-.-FTC), D-d4FC, emtricitabine,
phosphazide, fozivudine tidoxil, fosalvudine tidoxil, apricitibine
(AVX754), amdoxovir, KP-1461, abacavir+lamivudine,
abacavir+lamivudine+zidovudine, zidovudine+lamivudine,
[0183] 4) a HIV nucleotide inhibitor of reverse transcriptase,
e.g., tenofovir, tenofovir disoproxil fumarate+emtricitabine,
tenofovir disoproxil fumarate+emtricitabine+efavirenz, and
adefovir,
[0184] 5) a HIV integrase inhibitor, e.g., curcumin, derivatives of
curcumin, chicoric acid, derivatives of chicoric acid,
3,5-dicaffeoylquinic acid, derivatives of 3,5-dicaffeoylquinic
acid, aurintricarboxylic acid, derivatives of aurintricarboxylic
acid, caffeic acid phenethyl ester, derivatives of caffeic acid
phenethyl ester, tyrphostin, derivatives of tyrphostin, quercetin,
derivatives of quercetin, S-1360, zintevir (AR-177), L-870812, and
L-870810, MK-0518 (raltegravir), BMS-707035, MK-2048, BA-011,
BMS-538158, GSK364735C,
[0185] 6) a gp41 inhibitor, e.g., enfuvirtide, sifuvirtide, FB006M,
TRI-1144, SPC3, DES6, Locus gp41, CovX, and REP 9,
[0186] 7) a CXCR4 inhibitor, e.g., AMD-070,
[0187] 8) an entry inhibitor, e.g., SPO1A, TNX-355,
[0188] 9) a gp120 inhibitor, e.g., BMS-488043 and BlockAide/CR,
[0189] 10) a G6PD and NADH-oxidase inhibitor, e.g., immunitin, 10)
a CCR5 inhibitor, e.g., aplaviroc, vicriviroc, INCB9471, PRO-140,
INCB15050, PF-232798, CCR5mAb004, and maraviroc,
[0190] 11) an interferon, e.g., pegylated rIFN-alpha 2b, pegylated
rIFN-alpha 2a, rIFN-alpha 2b, IFN alpha-2b XL, rIFN-alpha 2a,
consensus IFN alpha, infergen, rebif, locteron, AVI-005,
PEG-infergen, pegylated IFN-beta, oral interferon alpha, feron,
reaferon, intermax alpha, r-IFN-beta, infergen+actimmune, IFN-omega
with DUROS, and albuferon,
[0191] 12) ribavirin analogs, e.g., rebetol, copegus, levovirin,
VX-497, and viramidine (taribavirin)
[0192] 13) NS5a inhibitors, e.g., A-831, A-689, and BMS-790052,
[0193] 14) NS5b polymerase inhibitors, e.g., NM-283,
valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB 1941,
MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433, and
XTL-2125,
[0194] 15) NS3 protease inhibitors, e.g., SCH-503034 (SCH-7),
VX-950 (Telaprevir), ITMN-191, and BILN-2065,
[0195] 16) alpha-glucosidase 1 inhibitors, e.g., MX-3253
(celgosivir) and UT-231B,
[0196] 17) hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and
LB-84451,
[0197] 18) non-nucleoside inhibitors of HCV, e.g., benzimidazole
derivatives, benzo-1,2,4-thiadiazine derivatives, and phenylalanine
derivatives,
[0198] 19) other drugs for treating Hepatitis C, e.g., zadaxin,
nitazoxanide (alinea), BIVN-401 (virostat), DEBIO-025, VGX-410C,
EMZ-702, AVI 4065, bavituximab, oglufanide, PYN-17, KPE02003002,
actilon (CPG-10101), KRN-7000, civacir, GI-5005, ANA-975
(isatoribine), XTL-6865, ANA 971, NOV-205, tarvacin, EHC-18, and
NIM811,
[0199] 20) pharmacokinetic enhancers, e.g., BAS-100 and SPI452, 20)
RNAse H inhibitors, e.g., ODN-93 and ODN-112, and
[0200] 21) other anti-HIV agents, e.g., VGV-1, PA-457 (bevirimat),
ampligen, HRG214, cytolin, polymun, VGX-410, KD247, AMZ 0026, CYT
99007, A-221 HIV, BAY 50-4798, MDX010 (iplimumab), PBS119, ALG889,
and PA-1050040.
[0201] In one embodiment, the additional therapeutic agent is
ribavirin. Accordingly, methods described herein include a method
of treating hepatitis C in a human patient in need thereof
comprising administering to the patient a therapeutically effective
amount of ribavirin and a therapeutically effective amount of a
pharmaceutical composition, pharmaceutical dosage form, or tablet
as described herein. In a further embodiment, the ribavirin and
pharmaceutical composition, pharmaceutical dosage form, or tablet
comprising sofosbuvir and ledipasvir is administered for about 12
weeks or less. In further embodiments, the ribavirin and
pharmaceutical composition, pharmaceutical dosage form, or tablet
comprising sofosbuvir and ledipasvir is administered for about 8
weeks or less, for about 6 weeks or less, or for about 4 weeks or
less.
[0202] It is contemplated that the additional therapeutic agent
will be administered in a manner that is known in the art and the
dosage may be selected by someone of skill in the art. For example,
the additional agent may be administered in a dose from about 0.01
milligrams to about 2 grams per day.
EXAMPLES
[0203] In the following examples and throughout this disclosure,
abbreviations as used herein have respective meanings as
follows:
TABLE-US-00001 ACN Acetonitrile AE Adverse Event API Active
Pharmaceutical Ingredient AUC Area Under the Curve AUC.sub.inf Area
under the concentration versus time curve extrapolated to infinite
time, calculated as AUC0 - last + (Clast/.lamda.z) AUC.sub.last
Area under the concentration versus time curve from time zero to
the last quantifiable concentration BMI Body Mass Indec BT
Breakthrough Rate CI Confidence Interval CL/F Apparent oral
clearance after administration of the drug: CL/F = Dose/AUC
C.sub.last Last observed quantifiable concentration of the drug cm
Centimeter C.sub.max Maximum Concentration cP Centipoise cP
Centipoise CV Coefficient of Variation D.sub.90 Particle Size DCF
Drug Content Factor DCF Drug Content Factor DCM Dichloromethane dL
Deciliter DRM Drug Related Material DSC Differential Scanning
Calorimetry E.sub.max Maximum Effect F % Percent Bioavailability
FaSSIF Fasted State Simulated Intestinal Fluids FB Free Base FDC
Fixed-Dose Combination FeSSIF Fed State Simulated Intestinal Fluid
FT Fourier Transform g Gram GLSM Geometric Least Squares Mean GMR
Geometric Mean Ratio GT Genotype h or hr Hour HCV Hepatitis C virus
HDPE High Density Polyethylene HPC Hydroxypropylcellulose HPLC
High-performance Liquid Chromatography HPMC Hydroxymethylcellulose
ICH International Conference on Harmonisation; Impurities
guidelines IFN Interferon IU International Unit KF Karl Fischer kg
Kilogram L Liter LCT Long Chain Triglycerides LDV Compound I,
GS-5885, Ledipasvir LLOD Lower limit of detection LLOQ Lower Limit
of Quantification LOD Limit of Detection M Molar mg Milligram min
Minute mL Milliliter mm Millimeter mM Millimolar N Population Size
n Number of Patients ng Nanogram nM Nanomolar nm Nanometer .degree.
C. Degrees Celsius PD Pharmacodynamic(s) PEG or PG Polyethylene
Glycol P-gp or Pgp P-glycoprotein PI Protease-Inhibitor PK
Pharmacokinetic PLS Partial Least Squares PPI Proton-Pump
Inhibitors PS Particle Size PVP Povidone PVP/VA Copovidone QS
Quantum Satis RAV Resistance Associated Variants RBV Ribavirin RH
Relative Humidity RNA Ribonucleic Acid RSD Relative Standard
Deviation RT Room Temperature S.sub.0 Intrinsic Solubility SAE
Serious Adverse Event SCT Short Chain Triglycerides SIBLM Simulated
Intestinal Bile Salt and Lecithin Mixture SIF Simulated Intestinal
Fluids SLS Sodium Lauryl Sulfate SOF Sofosbuvir (GS-7977, formerly
PSI-7977) SS-NMR Solid State Nuclear Magnetic Resonance SVR
Sustained Virologic Response t Time t.sub.1/2 Half-life (h) TFA
Trifluoroacetic acid T.sub.max Time (observed time point) of
C.sub.max UPLC Ultra Performance Liquid Chromatography Upper Resp
Upper Respiratory Tract Infection Tract Infx USP Uniform Standards
and Procedures UV Ultraviolet VL Viral Load vRVR Very Rapid Viral
Response V.sub.z/F Apparent volume of distribution wt or w Weight
XRPD Xray Powder Diffraction .mu.g Microgram .mu.L Microliter .mu.m
Micrometer
Example 1: Synthesis of Amorphous Ledipasvir
[0204] Methods for making various forms of ledipasvir may be found
in U.S. Publication Nos. 2013/0324740, and 2013/0324496. Both of
these applications are incorporated herein by reference. Following
is a method for isolating amorphous free base of ledipasvir.
[0205] Combine ledipasvir acetone solvate (191.4 g) and
acetonitrile (1356 g) in a reaction vessel and mix contents until a
solution is achieved. Add this ledipasvir in acetonitrile solution
slowly to another reaction vessel containing vigorously agitated
water (7870 g). Agitate contents at about 23.degree. C. for about
30 minutes. Filter the contents and dry at about 40-45.degree. C.
until constant weight is achieved to afford ledipasvir amorphous
solid (146.4 g, 82% yield).
Example 2: Tablet Preparation and Formulation
A. Dose Selection of Tablets
[0206] i. Sofosbuvir
[0207] The sofosbuvir dose selected for the tablet formulation is
400 mg once daily. Support for the 400 mg sofosbuvir dose can be
derived from E.sub.max PK/PD modeling with early virological and
human exposure data which also supports the selection of a 400 mg
sofosbuvir dose over others tested.
[0208] The mean sofosbuvir major metabolite AUC.sub.tau for the 400
mg sofosbuvir dose is associated with approximately 77% of the
maximal HCV RNA change from baseline achievable as determined by
this model, a value which is on the cusp of the plateau of the
exposure-response sigmoidal curve. In a sigmoidal E.sub.max model,
there is a relatively linear exposure-response relationship in the
20 to 80% maximal effect range. Therefore, given that sofosbuvir
exposure with 200 mg tablets appears dose-proportional with single
doses up to 1200 mg, doses below 400 mg are expected to yield
considerable reductions in the magnitude of HCV RNA change from
baseline. Similarly, in order to improve upon an efficacy
prediction of 77% in the plateau of the exposure-response curve,
substantial increases in exposure (and hence dose) would be needed
for an appreciable increase in antiviral effect.
[0209] The sofosbuvir dose of 400 mg once daily was associated with
higher SVR rates in genotype 1 HCV infected patients as compared to
the 200 mg once daily dose when given in conjunction with
additional HCV therapeutics for 24 weeks. Safety and tolerability
appeared similar across both dose levels. In addition, when
sofosbuvir 400 mg once daily plus other HCV therapeutics were given
to genotype 2 or 3 HCV infected patients, 100% SVR24 was
observed.
ii. Ledipasvir
[0210] The maximum median HCV RNA log 10 reduction was 3 or greater
for all cohorts dosed with .gtoreq.3 mg of ledipasvir. An E.sub.max
PK/PD model indicates that the exposures achieved following
administration of the 30 mg dose provides >95% of maximal
antiviral response in genotype 1a HCV infected patients. It was
also observed that 30 mg or greater of ledipasvir likely provided
coverage of some drug related mutations that doses less than 30 mg
did not, based on an analysis of NS5A mutants that arose in
response to exposure to ledipasvir. Therefore, 30 mg and 90 mg of
ledipasvir were selected as the dose for the formulations described
herein.
[0211] Further studies suggest that, when ledipasvir is
administered in combination with other therapeutic agents, the
breakthrough (BT) rate (number of patients with HCV RNA>lower
limit of quantification (LLOQ) after having achieved vRVR/total
number of patients who achieved vRVR), is higher with doses of 30
mg (BT=33%, 11/33; 30 mg ledipasvir), than with doses of 90 mg
(BT=12%, 9/74; 90 mg ledipasvir). Therefore, the 90 mg dose of
ledipasvir may confer a greater antiviral coverage that prevents
viral breakthrough.
B. Solid Dispersion Comprising Ledipasvir
[0212] To make the tablets comprising the combination of sofosbuvir
and ledipasvir as described herein, a solid dispersion comprising
ledipasvir was co-formulated with crystalline sofosbuvir. The
starting material of the solid dispersion can be a variety of forms
of ledipasvir including crystalline forms, amorphous form, salts
thereof, solvates and free base, as described herein. Because of
the high solubility in organic solvents and excipients and the
ability to isolate the ledipasvir free base crystalline acetone
solvate, this form was used in the amorphous solid dispersion of
ledipasvir.
[0213] The spray dried solid dispersion approach achieved the most
desirable characteristics relative to the other formulation
approaches, which included improved in vivo and in vitro
performance and manufacturability/scalability.
[0214] The spray dry feed solution was prepared by solubilizing
ledipasvir acetone solvate and polymer in the feed solvent.
Aggressive mixing or homogenization was used to avoid clumping of
the composition.
[0215] Different polymers were tested for preferred characteristics
in the solid dispersions. Non-ionic polymers such as hypromellose
and copovidone solid dispersions both showed adequate stability and
physical characteristics.
[0216] The feed solution was initially evaluated for appropriate
solvent with regard to solubility, stability, and viscosity.
Ethanol, methanol, and dichloromethane (DCM) all demonstrated
excellent solubility (ledipasvir solubility >500 mg/mL).
Ethanolic and DCM-based feed stocks were assessed for preparation
ease and spray dried at a range of inlet and outlet temperatures to
assess the robustness of the spray dry process. Both solvents gave
rapid dissolution of ledipasvir and copovidone.
[0217] Spray drying out of ethanol resulted in high yields (88, 90,
92, 94, 95, 97, 98, 99%) across a wide range of spray-drying outlet
temperatures (49-70.degree. C.) with no material accumulation on
the spray dry chamber. Spray drying out of DCM resulted in yields
of 60%, 78%, and 44%. Overall, the ledipasvir Solid Dispersion (50%
w/w) in a ledipasvir to copovidone ratio of 1:1 demonstrated good
chemical stability in the ethanolic feed solution.
[0218] An ethanolic solution of 10% ledipasvir acetone solvate and
10% copovidone was prepared using homogenization. Viscosity of
ethanolic solutions of ledipasvir:copovidone were low, measured
through 30% solids content (.about.65 cP).
[0219] Spray drying was conducted using two fluid nozzle or a
hydrolytic pressure nozzle. Table 1 presents the spray dry process
parameters evaluated at 100 g-4000 g of total feed solution using
the Anhydro MS35 spray dryer and Table 2 shows the spray dry
process parameters using the hydrolytic pressure nozzle. Particle
size data suggested sufficiently large particle size (10-14 .mu.m
mean PS) and was minimally affected by using higher spray rates or
a larger diameter spray nozzle. Nozzle gas flow was not modulated
to increase particle size.
TABLE-US-00002 TABLE 1 Ledipasvir Spray Dry Parameters on Anhydro
MS35 Spray Dryer Using a Two Fluid Nozzle Parameter Trial 1 Trial 2
Trial 3 Trial 4 Batch Size (g) 100 250 250 4000 Solids % 20 20 20
20 Feed Rate (mL/min) 30 40 40 40 Spray Nozzle (mm) 1.0 1.0 1.2 1.2
Nozzle Gas Flow (kg/hr) 6.0 6.0 6.0 6.0 Chamber Gas Flow 35.0 35.0
35.0 35.0 (kg/hr) Inlet Temp (.degree. C.) 125 165 165 165 Outlet
Temp (.degree. C.) 70 73 72 76 PS d.sub.10/d.sub.50/d.sub.90/mean
(.mu.m) 4/9/18/10 5/10/20/12 5/10/19/11 6/12/22/14 Post Spray LOD
(%) 5.56 4.86 4.29 3.42
TABLE-US-00003 TABLE 2 Example of Ledipasvir Spray Dry Parameters
Using a Hydrolyic Pressure Nozzle Parameter Trial 1 Batch Size (kg)
200 Solids % 20 Feed Rate (kg/hr) 178 Pressure Feed (bar) 52 Inlet
Temp (.degree. C.) 158 Outlet Temp (.degree. C.) 65 PS
d.sub.10/d.sub.50/d.sub.90/mean (.mu.m) 3/14/34 Post Spray LOD (%)
0.6
[0220] Organic volatile impurities, including the spray dry solvent
ethanol and residual acetone from ledipasvir acetone solvate are
rapidly removed during secondary drying at 60.degree. C. Smaller
scale production can be tray dried. On larger scale batches, a
double cone dryer or an agitated dryer can be used. Loss on drying
(LOD) was proportionately slower and is attributable to water,
which was later confirmed by Karl Fischer titration.
[0221] Residual ethanol was reduced below ICH guidelines of 0.5%
w/w by 6 hours of drying (or 8 hours for larger scale). Ethanol
content upon completion of drying was 0.08% w/w, and residual
acetone was 0.002%, indicating that the secondary drying process is
adequate for removal of residual solvent.
C. Tablet Preparation
[0222] i. Monolayer Tablet
[0223] Ledipasvir:copovidone solid dispersion (1:1) was made by
dissolving ledipasvir and copovidone into ethanol, and then spray
drying the mixture. The spray dried ledipasvir:copovidone solid
dispersion is further dried in a secondary dryer. The amorphous
solid dispersion comprising ledipasvir was blended with sofosbuvir
and excipients and milled to facilitate mixing and blend
uniformity. Either a coblend or codry granulation process can be
used. Coblend granulation is a multi-step process consisting of
separate dry granulations for each active ingredient with
excipients followed by the blending of the two granulations
together. Codry granulation consisted of dry granulating both
active ingredients and excipients together. The coblend and codry
processes demonstrated comparable physical and chemical tablet
properties. Exemplary coblend and codry formulations are provided
in Table 3 and Table 4 shown below.
TABLE-US-00004 TABLE 3 Representative Example Composition of
Sofosbuvir/Ledipasvir Codry (Co-granulated) Tablets at Various Fill
Weights % w/w Tablet Intra-granular Sofosbuvir 50.00 40.00 36.36
33.33 Ledipasvir:Copovidone 22.50 18.00 16.36 15.00 Solid
Dispersion (1:1) Lactose Monohydrate 6.67 16.33 23.19 26.11
Microcrystalline cellulose 3.33 8.17 11.60 13.05 Croscarmellose
Sodium 2.50 2.50 2.50 2.50 Silicon Dioxide 1.00 1.00 1.00 1.00
Magnesium stearate 0.75 0.75 0.75 0.75 Extra-granular
Microcrystalline cellulose 10.00 10.00 5.00 5.00 Croscarmellose
Sodium 2.50 2.50 2.50 2.50 Magnesium stearate 0.75 0.75 0.75 0.75
Fill wt (mg) 800 1000 1100 1200
TABLE-US-00005 TABLE 4 Representative Example Composition of
Sofosbuvir/Ledipasvir Coblend (Bi-granulated) Tablets % w/w
Intra-granular % w/w mg/ Composition Blend Tablet Tablet Sofosbuvir
Sofosbuvir 80 40 400 Intra-granular Microcrystalline 6 3 30 Blend
cellulose Lactose Monohydrate 6 3 30 Croscarmellose 4 2 20 Sodium
Silicon Dioxide 3 1.5 15 Magnesium stearate 1 0.5 5 Intra-granular
Subtotal 100 50 500 Ledipasvir Ledipasvir:Copovidone 42.4 18 180
Intra-granular Solid Dispersion Blend Microcrystalline 43.5 18.5
185 cellulose Croscarmellose 9.4 4 40 Sodium Silicon Dioxide 3.5
1.5 15 Magnesium stearate 1.2 0.5 5 Intra-granular Subtotal 100
42.5 425 Extra-granular Microcrystalline -- 5 50 cellulose
Croscarmellose -- 2 20 Sodium Magnesium stearate -- 0.5 5 Total 100
1000 Coating Film Coat -- 3 30 Purified water -- -- --
[0224] The granules were then mixed with a lubricant prior to
tablet compression. The total resulting core tablet weight was 1000
mg.
[0225] Film-coating of the tablets is provided to reduce photolytic
degradation. Tablets were coated to a target 3% weight gain. The
film-coating material was a polyvinylalcohol-based coating.
Exemplary tablet formulation is provided in Table 5.
TABLE-US-00006 TABLE 5 Representative Example of the Composition of
Tablets Comprising the Solid Dispersion of Ledipasvir and
Sofosbuvir Component Weight Ingredient % w/w (mg/tablet) Sofosbuvir
40.00 400 Ledipasvir Solid Dispersion 18.00 180.0 Lactose
Monohydrate 16.50 165.0 Microcrystalline Cellulose 18.00 180.0
Croscarmellose Sodium 5.00 50.0 Colloidal Silicon Dioxide 1.00 10.0
Magnesium Stearate 1.50 15 Total Tablet Core Weight 100.0 1000.0
Film coating 3.00 30.0 Purified Water -- -- Total Coated Tablet
Weight 1030.0
ii. Bilayer Tablet
[0226] Tablets comprising the co-formulation of a solid dispersion
comprising ledipasvir and crystalline sofosbuvir can also be made
as a bilayer tablet wherein each active ingredient is in a separate
layer. To make the bilayer tablet, a ledipasvir:copovidone (1:1)
solid dispersion is made by dissolving ledipasvir and copovidone
into ethanol, and then spray drying the mixture. The spray dried
ledipasvir:copovidone solid dispersion is further dried in a
secondary dryer. Next, the spray dried ledipasvir:copovidone solid
dispersion is then blended with excipients. The mixture is milled
and then blended with lubricant prior to dry granulation. The
ledipasvir granules are blended with extragranular lubricant.
Separately, the sofosbuvir drug substance is blended with
excipients, and then the mixture is milled and then blended with
lubricant prior to dry granulation. The sofosbuvir granules are
then blended with extragranular lubricant. Finally, the sofosbuvir
powder blend and ledipasvir powder blend are compressed into
bilayer tablet cores. The bilayer tablet cores are then film-coated
prior to packaging. A representative example composition of a
bilayer tablet comprising the solid dispersion of ledipasvir and
sofosbuvir is shown in Table 6. In this table, the solid dispersion
comprises ledipasvir:copovidone in a 1:1 ratio.
TABLE-US-00007 TABLE 6 Representative Example of Composition of
Bilayer Tablets Comprising the Solid Dispersion of Ledipasvir and
Sofosbuvir Component Weight Ingredient % w/w (mg/tablet) Layer 1
Sofosbuvir 33.34 400.0 Lactose Monohydrate 5.66 68.0
Microcrystalline Cellulose 7.50 90.0 Croscarmellose Sodium 2.00
24.0 Colloidal Silicon Dioxide 0.50 50.0 Magnesium Stearate 1.00
12.0 Layer 2 Ledipasvir Solid Dispersion 15.00 180.0 Lactose
Monohydrate 15.00 180.0 Microcrystalline Cellulose 17.00 204.0
Croscarmellose Sodium 2.50 30.0 Magnesium Stearate 0.50 6.0 Total
Tablet Core 100.00 1200
Example 3: PK, Stability and Dissolution Properties of Ledipasvir
Single-Agent Tablets and Ledipasvir/Sofosbuvir Tablets and
Reduction of Food-Effect and Effects of Gastric Acid
Suppressants
A. Ledipasvir Single-Agent Tablets Bioavailability
[0227] A series of in vivo experiments were conducted to evaluate
the potential benefit of the solid dispersion approach relative to
conventional formulations, as well as to optimize the solid
dispersion by identifying the most beneficial polymer type and
relative polymer concentration within the dispersion.
[0228] Equivalent bioavailability was achieved between formulations
comprising the free base amorphous form (4% w/w, 10 mg amorphous
free base tablet) and formulations comprising the D-tartrate salt
of ledipasvir (5.85% w/w, 10 mg D-tartrate salt tablet), both using
conventional formulations, in the pentagastrin pretreated dog
model, as shown in Table 7. Pentagastrin is a synthetic polypeptide
that stimulates the secretion of gastric acid, pepsin, and
intrinsic factor.
TABLE-US-00008 TABLE 7 Mean (RSD) Pharmacokinetic Parameters of
Ledipasvir Following Oral Administration of Tablets, 25 mg, in
Beagle Dogs (n = 6) AUC.sub.0-24 Drug Substance Form Pretreatment
C.sub.max (nM) (nM*hr) F (%) Amorphous Free base Pentagastrin 743
(17) 8028 (22) 71 Crystalline D-tartrate Pentagastrin 665 (38) 7623
(44) 67
[0229] Because these formulations displayed similar PK properties
and the isolation properties of the D-tartrate salt were preferable
to the free base amorphous form, the crystalline D-tartrate salt
formulation was chosen to compare to the amorphous solid dispersion
compositions. For these studies, 30 mg tablets comprising the
crystalline D-tartrate salt of ledipasvir and 30 mg or 90 mg
tablets comprising the amorphous solid dispersion of ledipasvir
were used. Dog pharmacokinetic results for select immediate release
ledipasvir tablets comprising ledipasvir solid dispersions are
shown in Table 8.
TABLE-US-00009 TABLE 8 Mean (RSD) Pharmacokinetic Parameters of
Ledipasvir after Oral Administration of Ledipasvir Tablets, Fasted
Beagle Dogs (n = 6) Ledipasvir: polymer Dose C.sub.max AUC.sub.0-24
F Polymer Ratio (mg) Pretreatment (nM) (nM*hr) (%) Crystalline N/A
30 Pentagastrin 665 7623 (44) 67 D-tartrate (38) Ledipasvir
Famotidine 154 1038 (41) 9 Tablets (44) 90 Pentagastrin 1831 18086
(36) 54 (28) Famotidine 349 3322 (40) 10 (37) Amorphous 2:1 30
Famotidine 251 2553 (54) 22 Solid (51) Dispersion Ledipasvir
Tablet: HPMC Amorphous 2:1 30 Famotidine 369 3383 (36) 30 Solid
(26) Dispersion 1:1 Pentagastrin 983 10541 (24) 93 Ledipasvir (22)
Tablet: 1:1 Famotidine 393 3930 (20) 35 Copovidone (30) 1:1 90
Pentagastrin 1644 20908 (41) 62 (38) 1:1 Famotidine 740 7722 (28)
23 (24)
[0230] Compared to the crystalline D-tartrate ledipasvir
formulations, the amorphous solid dispersion tablets displayed
higher bioavailability with lower variability. In pentagastrin
pretreated animals, an approximate 40% increase in exposure and a
2-fold decrease in variability were noted. More importantly in
famotidine pretreated animals, up to a 3.5-fold increase in
bioavailability was observed compared to the D-tartrate salt tablet
formulations.
[0231] A copovidone-based dispersion increased bioavailability more
than the equivalent hypromellose-based formulation (F=30% and 22%,
respectively) when spray dried at 2:1 API:polymer ratio.
Bioavailability of the copovidone-based formulation was further
enhanced by increasing the fraction of polymer to a 1:1 ratio,
resulting in a bioavailability of 35% in famotidine pretreated
dogs.
[0232] Because of the improved in vivo performance and acceptable
stability and physical properties, a 1:1 mixture of
ledipasvir:copovidone was chosen as the spray-dried material.
[0233] Formulations comprising the amorphous solid dispersions
proved to be advantageous over formulations comprising either the
amorphous free base or the D-tartrate salt. It was observed that
the bioavailability of amorphous free base formulations was similar
to D-tartrate salt formulations. Additional data showed a decrease
in bioavailability when ledipasvir was dosed with gastric acid
suppressing agents (famotidine), indicating an unfavorable
drug-drug interaction in free base amorphous and D-tartrate salt
formulations of ledipasvir. A solid dispersion using spray drying
with a hydrophilic polymer was identified to have acceptable
stability, physical characteristics, and in vivo performance. A
rapidly disintegrating tablet was developed using a dry granulation
process and commonly used excipients. A bioavailability study
comparing formulations comprising the D-tartrate salt with
formulations comprising the amorphous solid dispersion showed
improved biopharmaceutical performance and overcame much of the
negative drug-drug interactions with acid suppressive therapies
seen in the D-tartrate salt formulations.
B. Ledipasvir+Sofosbuvir Tablets Bioavailability
[0234] PK results for the combination of sofosbuvir with ledipasvir
(wherein the ledipasvir is in solid dispersion with copovidone in a
1:1) are shown in Table 9, and demonstrate lack of a significant
interaction between sofosbuvir and ledipasvir.
TABLE-US-00010 TABLE 9 Pharmacokinetic Data for Sofosbuvir and
Ledipasvir on Administration of Sofosbuvir and Ledipasvir Alone or
in Combination Sofosbuvir (n = 17) Sofosbuvir + % GMR Mean (% CV)
Sofosbuvir alone Ledipasvir (90% CI) AUC.sub.inf 794 (36.4) 1750
(27.8) 229 (191, 276) (ng hr/mL) AUC.sub.last 788 (36.6) 1740
(27.8) 230 (191, 277) (ng hr/mL) C.sub.max (ng/mL) 929 (52.3) 1870
(27.9) 221 (176, 278) Metabolite I (n = 17) Sofosbuvir + % GMR Mean
(% CV) Sofosbuvir alone Ledipasvir (90% CI) AUC.sub.inf 1110 (31.6)
1950 (22.8) 182 (157, 210) (ng hr/mL) AUC.sub.last 1060 (32.7) 1890
(22.8) 179 (155, 207) (ng hr/mL) C.sub.max (ng/mL) 312 (38.7) 553
(26.6) 182 (154, 216) Metabolite II (n = 17) Sofosbuvir + % GMR
Mean (% CV) Sofosbuvir alone Ledipasvir (90% CI) AUC.sub.inf 10900
(17.5) 13000 (16.7) 119 (113, 125) (ng hr/mL) AUC.sub.last 10200
(17.9) 12100 (15.5) 119 (113, 126) (ng hr/mL) C.sub.max (ng/mL)
1060 (17.3) 864 (20.1) 81.2 (76.9, 85.8) Ledipasvir (n = 17)
Sofosbuvir + % GMR Mean (% CV) Ledipasvir alone Ledipasvir (90% CI)
AUC.sub.inf 11900 (26.2) 11400 (27.0) 95.7 (92.1, 99.5) (ng hr/mL)
AUC.sub.last 755 (24.7) 734 (27.0) 96.5 (89.9, 104) (ng hr/mL)
C.sub.max (ng/mL) 375 (28.8) 360 (31.2) 95.5 (91.9, 99.1)
[0235] Sofosbuvir plasma exposure was increased by .about.2.3-fold
by ledipasvir. The effect of ledipasvir on sofosbuvir is likely due
to inhibition of P-gp, of which Sofosbuvir is a known substrate.
The increase in sofosbuvir was not considered significant due to
its very low and transient exposure relative to total drug related
material (DRM) exposure (DRM, calculated as the sum of the AUCs for
each of the analytes, corrected for molecular weight). Based on
this calculation, the AUC of sofosbuvir with ledipasvir is only
.about.5.7% of DRM AUC. The exposure of metabolite II, the major
circulating sofosbuvir metabolite, was not impacted by the
administration of ledipasvir, and demonstrates the lack of
significant interaction between sofosbuvir and ledipasvir.
C. Reduction of Food Effect in Solid Dispersions of Ledipasvir and
Ledipasvir/Sofosbuvir Tablets
[0236] Ledipasvir alone in a conventional formulation (not the
solid dispersion) has been demonstrated to have a negative food
effect. Table 10 summarizes PK parameters of ledipasvir following a
single dose of ledipasvir, 30 mg, under fasted and fed conditions.
The ledipasvir PK profile was altered in the presence of food.
Specifically, the high-fat meal appeared to delay ledipasvir
absorption, prolong T.sub.max (median T.sub.max of 8 hours), and
decreased ledipasvir plasma exposure (approximately 45% decrease
each in mean C.sub.max, AUC.sub.last, and AUC.sub.inf,
respectively).
TABLE-US-00011 TABLE 10 Plasma Ledipasvir PK Parameters Following
Single-dose Administration of Ledipasvir by Concomitant Food Intake
Status Mean (% CV) Ledipasvir Ledipasvir 30 mg 30 mg Fed PK
Parameter (N = 8) (N = 8) C.sub.max (ng/mL) 73.1 (50.8) 36.5 (22.6)
T.sub.max (h) 6.00 (5.00, 6.00) 8.00 (7.00, 8.00) AUC.sub.last
1988.2 (58.2) 996.5 (21.6) (ng h/mL) AUC.sub.inf 2415.9 (60.3)
1175.0 (25.3) (ng h/mL) t.sub.1/2 (h) 39.82 (33.15, 41.65) 36.83
(22.19, 49.08) CL/F (mL/h) 17,034.5 (58.6) 26,917.9 (23.6)
V.sub.z/F (mL) 876,546.3 (44.2) 1,386,469 (24.9) C.sub.last (ng/mL)
6.8 (68.0) 3.1 (42.2)
[0237] Table 11 presents the ratio of the GLSMs (ledipasvir 30 mg
under fasted conditions/ledipasvir 30 mg under fed conditions) for
each of the primary PK parameters.
TABLE-US-00012 TABLE 11 Statistical Evaluations of Ledipasvir PK
Parameters for Food Effect Geometric Least Squares Mean (GLSM)
Ledipasvir Ledipasvir GLSM Ratio 90% 30 mg 30 mg (Fed/Fasted)
Confidence Fed (N = 8) Fasted (N = 8) % Interval C.sub.max (ng/mL)
35.87 65.33 54.90 39.10, 77.08 AUC.sub.last 977.76 1724.28 56.71
38.87, 82.73 (ng hr/mL) AUC.sub.inf 1143.64 2058.78 55.55 36.88,
83.67 (ng hr/mL)
[0238] Similar median half-lives of ledipasvir were observed
independent of administration under fasted or fed conditions
(t.sub.1/2 of 39.82 hours under fasted conditions vs 36.83 hours
under fed conditions) indicating that food decreased the
bioavailability of ledipasvir by reducing its solubility and/or
absorption.
[0239] Because ledipasvir has been demonstrated to have a negative
food effect, the composition comprising both sofosbuvir and
ledipasvir (as solid dispersion in copovidone (1:1)) was tested for
a food effect. These results are shown in Table 12. Food slowed the
rate of absorption of sofosbuvir (median T.sub.max: 1.00 vs 2.00
hours) with only modest alteration in the bioavailability, as
evidenced by increases of 2-fold or less in sofosbuvir and
sofosbuvir metabolite I plasma exposure. For sofosbuvir metabolite
II, an approximately 20-30% lower C.sub.max was observed upon
sofosbuvir administration with food with no change in AUC. The %
GMR and associated 90% CI (fed/fasted treatments) for AUC of
sofosbuvir metabolite II were within the equivalence bounds of 70%
to 143%. Since the decrease in sofosbuvir metabolite II C.sub.max
was modest and the AUC parameters met the equivalence criteria, the
effect of food on sofosbuvir metabolite II was not considered
significant.
[0240] Similar ledipasvir plasma exposures (AUC and C.sub.max) were
achieved upon administration of ledipasvir under fasted or fed
conditions. The % GMR and associated 90% CIs (fed/fasted
treatments) were within the equivalence bounds of 70-143%. While a
"negative" food effect was previously observed on ledipasvir when
administered alone (as the amorphous free base, not solid
dispersion), the pharmacokinetics of ledipasvir (amorphous solid
dispersion; copovidone (1:1)) administered in combination with
sofosbuvir does not appear to be altered by food. As such, the
combination of sofosbuvir and ledipasvir may be administered
without regard to food.
TABLE-US-00013 TABLE 12 Pharmacokinetic Data for Sofosbuvir,
Sofosbuvir Metabolites I and II, and Ledipasvir on Administration
of Sofosbuvir/Ledipasvir Tablets Fasted or with a Moderate-Fat Meal
or with a High-Calorie/High Fat Meal Sofosbuvir (n = 29)
Sofosbuvir/Ledipasvir Sofosbuvir/Ledipasvir Tablet Tablet % GMR
(90% CI) Mean (% CV) Fasted Moderate-Fat Meal [Moderate-Fat/Fasted]
AUC.sub.inf (ng hr/mL) 1520 (39.5) 2860 (33.4) 195 (176, 216)
AUC.sub.last (ng hr/mL) 1520 (39.7) 2850 (33.5) 195 (176, 216)
C.sub.max (ng/mL) 1240 (49.6) 1520 (39.8) 126 (109, 147)
Sofosbuvir/Ledipasvir Sofosbuvir/Ledipasvir Tablet Tablet
High-Calorie/High-Fat GMR (90% CI) Fasted Meal [High-Fat/Fasted]
AUC.sub.inf (ng hr/mL) 1520 (39.5) 2570 (34.0) 179 (162, 198)
AUC.sub.last (ng hr/mL) 1520 (39.7) 2550 (34.6) 178 (161, 198)
C.sub.max (ng/mL) 1240 (49.6) 1350 (42.5) 115 (99.0, 134)
Sofosbuvir Metabolite I (n = 29) Sofosbuvir/Ledipasvir
Sofosbuvir/Ledipasvir Tablet Tablet % GMR (90% CI) Mean (% CV)
Fasted Moderate-Fat Meal [Moderate-Fat/Fasted] AUC.sub.inf (ng
hr/mL) 1520 (42.0) 2520 (21.4) 177 (163, 192) AUC.sub.last (ng
hr/mL) 1470 (43.3) 2460 (21.8) 180 (164, 196) C.sub.max (ng/mL) 352
(42.7) 495 (22.2) 151 (136, 167) Sofosbuvir/Ledipasvir
Sofosbuvir/Ledipasvir Tablet Tablet High-Calorie/High-Fat GMR (90%
CI) Fasted Meal [High-Fat/Fasted] AUC.sub.inf (ng hr/mL) 1520
(42.0) 2550 (22.2) 181 (166, 196) AUC.sub.last (ng hr/mL) 1470
(43.3) 2500 (22.5) 184 (168, 201) C.sub.max (ng/mL) 352 (42.7) 501
(26.8) 154 (139, 171) Sofosbuvir Metabolite II (n = 29)
Sofosbuvir/Ledipasvir Sofosbuvir/Ledipasvir Tablet Tablet % GMR
(90% CI) Mean (% CV) Fasted Moderate-Fat Meal [Moderate-Fat/Fasted]
AUC.sub.inf (ng hr/mL) 11800 (23.0) 13800 (17.7) 117 (112, 123)
AUC.sub.last (ng hr/mL) 11300 (23.4) 12900 (18.2) 114 (108, 121)
C.sub.max (ng/mL) 865 (26.6) 700 (19.5) 81.5 (75.6, 87.9)
Sofosbuvir/Ledipasvir Sofosbuvir/Ledipasvir Tablet Tablet
High-Calorie/High-Fat GMR (90% CI) Fasted Meal [High-Fat/Fasted]
AUC.sub.inf (ng hr/mL) 11800 (23.0) 12900 (18.5) 112 (107, 118)
AUC.sub.last (ng hr/mL) 11300 (23.4) 12100 (20.1) 110 (103, 116)
C.sub.max (ng/mL) 865 (26.6) 600 (22.9) 70.2 (65.0, 75.8)
Ledipasvir (n = 29) Sofosbuvir/Ledipasvir Sofosbuvir/Ledipasvir
Tablet Tablet % GMR (90% CI) Mean (% CV) Fasted Moderate-Fat Meal
[Moderate-Fat/Fasted] AUC.sub.inf (ng hr/mL) 10600 (57.2) 10600
(35.6) 115 (99.4, 134) AUC.sub.last (ng hr/mL) 8600 (53.8) 8650
(32.1) 114 (98.0, 133) C.sub.max (ng/mL) 324 (44.8) 319 (24.8) 109
(93.5, 126) Sofosbuvir/Ledipasvir Sofosbuvir/Ledipasvir Tablet
Tablet High-Calorie/High-Fat GMR (90% CI) Fasted Meal
[High-Fat/Fasted] AUC.sub.inf (ng hr/mL) 10600 (57.2) 9220 (36.1)
103 (88.5, 119) AUC.sub.last (ng hr/mL) 8600 (53.8) 7550 (33.9) 104
(88.8, 121) C.sub.max (ng/mL) 324 (44.8) 255 (25.9) 88.2 (75.8,
103)
D. Reduction of Effects of Gastric Acid Suppressants in
Ledipasvir/Sofosbuvir Tablets
[0241] Ledipasvir, 30 mg, alone in both a conventional formulation
(as the D-tartrate salt) and as the solid dispersion has been
demonstrated to have a decrease in bioavailability when
administered with some gastric acid suppressants; most
significantly, proton-pump inhibitors (PPI's, e.g., omeprazole),
but also including histamine-2 antagonsists (H2RA's, e.g.,
famotidine, data not included). Table 12A summarizes PK parameters
of ledipasvir following administration of ledipasvir conventional
single agent tablets, 30 mg, ledipasvir tablets as solid dispersion
(ledipasvir:copovidone 1:1), 30 mg, and sofosbuvir/ledipasvir FDC
tablets (90 mg of ledipasvir solid dispersion comprising copovidone
1:1) with and without omeprazole. The bioavailability of ledipasvir
as single agent tablets was reduced approximately 2-fold when
administered with omeprazole; however, administration of ledipasvir
as part of the sofosbuvir/ledipasvir FDC tablet with omeprazole
resulted in no significant decrease in ledipasvir exposure (AUC and
C.sub.max) compared to sofosbuvir/ledipasvir FDC tablet
administration in absence of omeprazole.
TABLE-US-00014 TABLE 12A Pharmacokinetic Data for Ledipasvir on
Administration of Ledipasvir Single Agent Tablets or
Sofosbuvir/Ledipasvir Tablets with and without Omeprazole
Ledipasvir, Conventional Formulation (N = 10) Ledipasvir Ledipasvir
+ % GMR Mean (% CV) alone Omeprazole (90% CI) AUC.sub.tau (ng
hr/mL) 1640 (18.5) 865 (37.7) 50.7 (43.4, 59.3) C.sub.max (ng/mL)
99.0 (20.1) 51.2 (39.2) 49.7 (41.7, 59.1) C.sub.tau (ng/mL) 52.2
(22.1) 28.3 (36.0) 52.4 (44.3, 61.9) Ledipasvir, Solid Dispersion
(N = 17) Ledipasvir Ledipasvir + % GMR Mean (% CV) alone Omeprazole
(90% CI) AUC.sub.inf (ng hr/mL) 2140 (38.8) 1300 (50.7) 58.5 (48.3,
70.8) AUC.sub.last (ng hr/mL) 1850 (33.5) 1070 (45.5) 56.3 (46.4,
68.3) C.sub.max (ng/mL) 64.8 (32.9) 36.2 (55.9) 52.2 (41.4, 65.9)
Ledipasvir, SOF/LDV FDC (N = 16) SOF/LDV SOF/LDV FDC + % GMR Mean
(% CV) FDC Alone Omeprazole (90% CI) AUC.sub.inf (ng hr/mL) 7990
(66.2) 6660 (51.8) 96.0 (66.5, 139) AUC.sub.last (ng hr/mL) 7160
(65.8) 5700 (51.8) 92.5 (64.8, 132) C.sub.max (ng/mL) 242 (68.6)
176 (51.1) 89.1 (60.9, 130)
E. Dissolution of Ledipasvir/Sofosbuvir Tablets
[0242] Dissolution studies were conducted comparing the sofosbuvir
400 mg/ledipasvir 90 mg tablets (ledipasvir:copovidone (1:1). The
sofosbuvir/ledipasvir tablets (LOT 1-5) display greater than 85%
sofosbuvir (FIG. 5) and ledipasvir (FIG. 6) dissolved in 30 minutes
for both tablet formulations. These results are shown in FIGS. 5
and 6.
Example 4: Stability of Sofosbuvir/Ledipasvir Co-Formulation
[0243] The compatibility of sofosbuvir anhydrous crystalline drug
substance was evaluated with the ledipasvir:copovidone solid
dispersion. A blend of the sofosbuvir and ledipasvir:copovidone
(1:1) solid dispersion was prepared at a ratio representative of
the final 400 mg sofosbuvir/90 mg ledipasvir tablets. The blend was
compressed into pellets and placed in stability chambers at
40.degree. C./75% RH and 60.degree. C./ambient humidity and tested
after two and four weeks of storage in open glass vials. The
results summarized in Table 13 show that no degradation was
observed for either sofosbuvir or ledipasvir, demonstrating the
chemical compatibility of sofosbuvir and the ledipasvir:copovidone
solid dispersion with each other.
TABLE-US-00015 TABLE 13 Strength and Impurity Content of Sofosbuvir
and Ledipasvir:Copovidone Solid Dispersion Blend Stored at
40.degree. C./75% RH and 60.degree. C. Ledipasvir Sofosbuvir Total
Total Time Strength Impurity Strength Impurity Condition (weeks)
(%) Content (%) (%) Content (%) 45.degree. C./ 0 98.8 0.0 102.9 0.4
75% RH 2 96.9 0.0 101.6 0.3 4 97.1 0.0 100.5 0.2 60.degree. C. 0
98.8 0.0 102.9 0.4 1 99.2 0.0 102.4 0.3 2 99.6 0.0 103.2 0.3 4 98.9
0.0 102.8 0.2
Example 5: Efficacy of Sofosbuvir/Ledipasvir/Ribavirin Treatment in
Patients with HCV Infections
[0244] Patients with HCV infections were treated with either the
combination of sofosbuvir, ledipasvir, and ribavirin or the
combination of sofosbuvir and ribavirin. Patients used in the study
included those that were treatment naive, i.e. had not previously
been treated for HCV and those that were null responders, i.e. had
previously been treated for HCV but failed to respond to the
treatment. Standard doses (90 mg of ledipasvir, 400 mg of
sofosbuvir, and 1000 mg of ribavirin, for example) were given of
each drug to the patients for a duration of 12 weeks. Throughout
treatment, HCV RNA was measured, and the Sustained Virologic
Response (SVR) was measured after treatment was discontinued. By
four weeks of treatment, almost all patients had achieved an HCV
RNA measurement below the limit of detection (LOD of 15 IU/mL), and
by the end of treatment, 100% of patients achieved an HCV RNA level
below the LOD (Table 14).
TABLE-US-00016 TABLE 14 Patients with HCV RNA below the limit of
detection over time. Sofosbuvir + Ribavirin Sofosbuvir + Ribavirin
+ Ledipasvir Treatment-naive Null responder Treatment-naive Null
responder (n = 25) (n = 10) (n = 25) (n = 9) Week 1 32% 10% 44% 0%
Week 2 68% 70% 88% 44% Week 4 100% 100% 100% 89% End of Treatment
100% 100% 100% 100%
[0245] Surprisingly, 100% of patients receiving the combination of
sofosbuvir, ledipasvir, and ribavirin achieved a sustained
virologic response at four and twelve weeks post treatment. In
contrast, only 88% of treatment naive and 10% of null responder
patients treated with the combination of sofosbuvir and ribavirin
achieved a SVR at four weekes post treatment, and only 84% of
treatment naive and 10% of null responder patients treated with the
combination of sofosbuvir and ribavirin achieved SVR at twelve
weekes post treatment (Table 15).
TABLE-US-00017 TABLE 15 Sustained Virologic Response Sofosbuvir +
Ribavirin + Sofosbuvir + Ribavirin Ledipasvir Treatment- Null
responder Treatment- Null responder naive (n = 25) (n = 10) naive
(n = 25) (n = 9) SVR4 88% 10% 100% 100% SVR12 84% 10% 100% 100%
[0246] These results are graphically depicted as FIG. 7A-D and
demonstrate that the addition of ledipasvir in the treatment
regimen gave 100% SVR at weeks 4 and 12. Example 9, below, shows
similar results are obtained with treatment regimens of less than
twelve weeks (i.e. treatment regimens of about 8 or 6 weeks), and
that similar results are obtained with treatment regimens of
sofosbuvir and ledipasvir without the addition of ribavirin.
Example 6. Stability of SOF 400 mg/Ledipasvir 90 mg Fixed-Dose
Combination Tablets
[0247] This example summarizes the physicochemical stability of
packaged Sofosbuvir (SOF) 400 mg/ledipasvir 90 mg blue film-coated
fixed-dose combination (FDC) tablets at 25.degree. C./60% relative
humidity (RH) and 40.degree. C./75% RH as a function of desiccant.
The ledipasvir portion of the table comprised ledipasvir:copovidone
in a 1:1 ratio. In addition the chemical and physical stability of
SOF/ledipasvir FDC tablets were evaluated at 40.degree. C./75% RH
under open condition for up to 4 weeks.
[0248] The physico-chemical properties that were evaluated included
appearance, potency, degradant formation, dissolution rate and
water content. Physical stability of the tablets in the absence of
desiccant was evaluated after 24 weeks using FT-Raman spectroscopy
and modulated differential scanning calorimetry (mDSC).
[0249] SOF 400 mg/ledipasvir 90 mg blue film-coated FDC tablets
exhibited satisfactory stability at 25.degree. C./60% RH and
40.degree. C./75% RH for up to 24 weeks in the presence of 0, 1,
and 3 g of desiccant. No significant changes were observed in
potency, impurity content or dissolution rate. However, a
ledipasvir photodegradant was present at 0.1% for all conditions.
FT-Raman analysis for the tablets stored in the absence of
desiccant showed no detectable crystallization after 24-weeks.
Methods and Materials
Materials
[0250] Table 16 lists the physicochemical properties for SOF drug
substance and ledipasvir solid dispersion used to produce tablets.
The quantities of SOF drug substance and ledipasvir solid
dispersion were adjusted based on their respective drug content
factor (DCF) with concomitant adjustment in the quantity of lactose
monohydrate. The DCF used for SOF and ledipasvir solid dispersion
powder, 50% w/w were 0.997 and 0.497 (0.994 when adjusted for the
amount of copovidone), respectively.
TABLE-US-00018 TABLE 16 Physicochemical Properties of SOF Drug
Substance and Ledipasvir Solid Dispersion, 50% w/w, Bulk Powder
Used to Produce SOF 400 mg/ Ledipasvir 90 mg Film-Coated FDC
Tablets Water Assay by Drug Content by Particle Size Active Crystal
HPLC Impurities Content Karl Fischer (.mu.m) Ingredient Form (%)
(%) Factor (%) d.sub.10 d.sub.50 d.sub.90 SOF Anhydrous 99.8 0.1
0.996 0.1 3 10 29 Form II Ledipasvir Solid Amorphous 49.7 0.2 0.497
1.09 5 22 44 Dispersion, 50% w/w, Bulk Powder
Equipment
[0251] The primary equipment used to manufacture SOF 400
mg/Ledipasvir 90 mg film-coated FDC tablets included an 12 qt.
V-Blender, a screening mill (Comil 197S, Quadro, Waterloo, Canada)
equipped with a 0.094 in grated screen, a roller
compactor/granulator (MiniPactor, Gerteis, Jona, Switzerland)
equipped equipped with a 1.0 mm milling screen and a smooth/smooth
roller configuration, a 12-station instrumented rotary tablet press
(XM-12, Korsch, Berlin, Germany), and a tablet coater (LabCoat,
O'Hara Technologies Inc., Ontario, Canada). The diamond-shaped
tablet tooling (Elizabeth Carbide Die Co., Inc., McKeesport, Pa.,
USA) consisted of diamond, standard concave D-type punches with
dimensions of 0.7650 in x 0.4014 in (19.43 mm.times.10.20 mm). A 15
inch perforated pan film coater was used to coat the tablet
cores.
Container Closure
[0252] Sofosbuvir/Ledipasvir FDC tablets are packaged in 100 mL
white, high density polyethylene (HDPE) bottles. Each bottle
contained 30 tablets and 0, 1 or 3 g silica gel desiccant canister
or sachet and polyester packing material. Each bottle was enclosed
with a white, continuous thread, child-resistant screw cap with an
induction-sealed, aluminum-faced liner.
[0253] A selected number of bottles were left open and packaged
without desiccant to evaluate the physical and chemical stability
at 40.degree. C./75% RH under accelerated heat and humidity
conditions.
General Study Design
[0254] The solid state and chemical stability of the packaged lot
were evaluated in the following configurations:
1) At 25.degree. C./60% RH and 40.degree. C./75% RH as a function
of desiccant. The samples were stored under closed condition for a
minimum of 24 weeks. 2) At 40.degree. C./75% RH under open
condition for up to 4 weeks.
[0255] Samples were pulled at predetermined time points. Chemical
stability testing for appearance, potency, degradant formation,
dissolution rate and water content was conducted. Additional
physical stability assays to monitor potential crystallization and
phase separation were conducted.
Physical Stability Evaluation
[0256] Physical stability tests included appearance and FT-Raman.
The visual inspection was performed on stressed film-coated tablets
to identify changes in tablet color and coating integrity. FT-Raman
spectroscopy was used to detect potential crystalline ledipasvir
(Form III) in the film-coated tablets.
[0257] Tablets were visually inspected for changes in appearance at
all time points and storage conditions. In contrast, FT-Raman was
only performed on tablets with 0 g desiccant at 24 weeks
(25.degree. C./60% RH and 40.degree. C./75% RH).
Appearance
[0258] At all time points tablets were examined for physical
integrity (i.e. color, shape, coating integrity and debossing).
FT-Raman
[0259] FT-Raman experiments were conducted. The 24-week
SOF/ledipasvir film-coated FDC tablets stored in closed containers
at 25.degree. C./60% RH and 40.degree. C./75% RH were analyzed
using FT-Raman spectroscopy to detect the formation of crystalline
ledipasvir (Form III). Briefly, the coating from the tablets was
carefully removed using an Xacto.TM. knife followed by grinding of
the tablet in a mortar and pestle. Tablet powder was then packed
into cups and spectra were collected using a backscattering
geometry.
Chemical Stability Evaluation
[0260] Chemical stability assays included measuring water content
by Karl Fischer (KF), potency, formation of impurity/degradation
products and dissolution rate were conduced.
KF Water Content
[0261] The water content was reported for SOF 400 mg/ledipasvir 90
mg film-coated FDC tablets following USP <921>.
Potency and Impurity/Degradant Formation by UPLC
[0262] The potency and degradation product formation of
SOF/ledipasvir film-coated FDC tablets were evaluated by analysis
of composite sample solution of 10 tablets according to STM-2542
[5]. The reference standard concentration for SOF and ledipasvir is
2.0 mg/mL and 0.45 mg/mL, respectively. The strength and
degradation product content of SOF and ledipasvir was determined by
UPLC using external reference standard and area normalization at
wavelengths of 262 nm and 325 nm, respectively.
Dissolution Methodology
[0263] Dissolution testing was performed on SOF/ledipasvir
film-coated FDC tablets. A USP type 2 dissolution apparatus with
900 mL of dissolution medium and a paddle speed of 75 rpm was used.
The medium was 1.5% polysorbate 80 in 10 mM potassium phosphate
buffer at pH 6.0 and the temperature was maintained at 37.degree.
C. for the duration of the assay. The extent of SOF and ledipasvir
released as a function of time was monitored by UPLC using area
normalization and an external reference standard at a wavelength of
250 nm.
Results
A. Physical Stability
A1. Appearance
[0264] Samples at all stability conditions and desiccant levels
were visually inspected for all time points and found to resemble
blue, diamond-shaped film-coated tablets.
A2. FT-Raman
[0265] The FT-Raman analysis was performed on powder extracted from
tablets stored in the absence of desiccant after 24 weeks.
Calculations of % crystallinity using the PLS model did not show
signs of crystalline ledipasvir (Form III) above the LOD of 3% at
either storage condition. This was consistent with the original
sample (t=0) in which ledipasvir (Form III) was also below the LOD.
Spectra from selected samples were included in a chart, from 1577
cm.sup.-1 to 1514 cm.sup.-1 with the baselines artificially
adjusted for clarity. This region is in one of the four spectral
regions used to estimate the % ledipasvir (Form III) in tablets by
PLS model.
[0266] The top two spectra (used as standards in the PLS model), in
the chart, were from tablets spiked with 10% w/w and 3% w/w, of
crystalline ledipasvir (Form III). The next two spectra represent
stressed tablets stored for 24 weeks at 40.degree. C./75% RH and
25.degree. C./60% RH. The last spectrum represents the initial time
point (t=0). Ledipasvir (Form III) has a distinct peak at 1552
cm.sup.-1, which can clearly be seen in the spiked tablets with
increasing intensity from 3% to 10%. The intensity in this region
for the stressed samples stored for 24 weeks does not increase from
the t=0 sample, indicating no change in crystallinity. Ledipasvir
(Form III) in the t=0 sample and the 24 week samples is below that
present in the tablets spiked with 3% Form III ledipasvir, the
current limit of detection for this analytical technique.
B. Chemical Stability
B.1 KF Water Content
[0267] The water content of stressed samples stored for 4 weeks
under open condition increased from 2.28% to 5.23%. The amount of
water content of stressed samples stored at 25.degree. C./60% RH
decreased to 1.91%, 1.58%, and 1.65% for tablets with no desiccant,
with 1 g desiccant, and 3 g desiccant, respectively. At 40.degree.
C./75% RH, the amount of water content decreased to 2.03%, 1.79%,
and 1.46% for tablets without desiccant, with 1 g desiccant, and 3
g desiccant, respectively.
B.2 Potency and Impurity/Degradation Product Formation
[0268] The potency and impurity/degradation content for SOF 400
mg/ledipasvir 90 mg film-coated FDC tablets were determined at
25.degree. C./60% RH and 40.degree. C./75% RH. Representative
chromatograms of stability samples stored at 40.degree. C./75% RH
were obtained. The data showed that SOF and ledipasvir remained
chemically stable in SOF 400 mg/ledipasvir 90 mg film-coated FDC
tablets stored for 24 weeks at 25.degree. C./60% RH and 40.degree.
C./75% RH. The label strength for SOF and ledipasvir remains
unchanged at 25.degree. C./60% RH and 40.degree. C./75% RH.
Dissolution
[0269] The dissolution profiles of SOF and ledipasvir in SOF 400
mg/ledipasvir 90 mg film-coated FDC tablets were obtained. At the
24 week time point, the tablets ranged between 99% and 100%
dissolution at 45 minutes for SOF, and between 99% and 98% for
ledipasvir at both 25.degree. C./60% RH and 40.degree. C./75% RH
for all desiccant levels tested.
[0270] From the foregoing, this example shows that SOF 400
mg/ledipasvir 90 mg Film-Coated FDC tablets exhibited satisfactory
stability at 25.degree. C./60% RH and 40.degree. C./75% RH for up
to 24 weeks in the presence of 0, 1, and 3 g of desiccant. In
addition, crystalline ledipasvir (Form III) was not detected by
FT-Raman analysis after 24 weeks of storage.
Example 7. Formulation Development of a Fixed Dose Combination
(FDC) Tablet SOF 400 mg/Ledipasvir 90 mg
[0271] This example shows the development of a SOF 400
mg/ledipasvir 90 mg fixed dose combination (FDC) tablet comprising
ledipasvir:copovidone (1:1). There were expected difficulties with
such a development, one of which was the expected poor powder flow
and the other relates to non-homogenous blend, given the existing
formulations of each individual agent.
[0272] Three tablet formulations were tested, including (1) a
monolayer co-granulated tablet formulation, (2) a monolayer
co-blended tablet formulation and (3) a bilayer tablet formulation.
In all of these formulations, SOF was in anhydrous crystalline form
II and ledipasvir was in amorphous solid dispersion
(ledipasvir:copovidone (1:1)).
[0273] Formulation (1) is typically associated with the highest
risk of drug-drug interaction but is the most cost-effective during
manufacturing. The bilayer formuation of (3), by constrast, is
perceived to have the lowest drug-drug interaction risk.
[0274] The dissolution performance of the formulations were tested
in a dissolution media that included 10 mM phosphate buffer at pH
6.0 (1.5% Tween.RTM. 80). As shown in FIG. 8A-B, all three
formulations had comparable dissolution performance, similar to
that of the single-agent controls.
[0275] The pharmacokinetic (PK) performance of each formulation was
also tested. Plasma Concentration of SOF/ledipasvir after oral
administration of SOF/ledipasvir FDC and control tablets in fasted
dogs (100 mg/22.5 mg fixed/dog). Table 17 below shows the PK
results.
TABLE-US-00019 TABLE 17 Phamacokinetic performance of the
forumations in famotidine pretreated dogs Total Tablet SOF
Ledipasvir weight/ AUC.sub.0-t C.sub.max AUC.sub.0-last C.sub.max
Formulation Treatment (ng*hr/mL) (ng/mL) (ng*hr/mL) (ng/mL) Control
Famotidine 314 .+-. 207 503 .+-. 363 3260 .+-. 1312 345 .+-. 132
SOF tablet + Ledipasvir SD tablet Monolayer, Famotidine 501 .+-.
249 729 .+-. 434 3236 .+-. 730 333 .+-. 56 Co-granulated Monolayer,
Famotidine 483 .+-. 406 652 .+-. 527 4208 .+-. 2216 444 .+-. 215
Co-blended Bilayer Famotidine 283 .+-. 193 288 .+-. 201 4,712 .+-.
2,270 421.7 .+-. 203.7
[0276] Based on these results, the monolayer co-granulated tablet
was selected for further analysis. The composition of this
formulation is provided in Table 18.
TABLE-US-00020 TABLE 18 Composition of SOF 400 mg/Ledipasvir 90 mg
FDC Tablets Composition % w/w Intra-granular SOF 40.00% Ledipasvir
SD 18.00% Lactose Fast Flow 316 16.50% MCC 101 8.00% Croscarmellose
2.50% Silicon Dioxide 1.00% Magnesium Stearate 0.75% Extragranular
MCC 101 10.00% Croscarmellose 2.50% Magnesium stearate 0.75% Total
Fill weight Core Tablet 1000 (mg) Coating Opadry II Orange 85F13912
3.0% Water QS
[0277] A bioavailability clinical study was carried out with this
formulation, with single agent tablets as control, in 24 healthy
patients under fasted conditions. The results are shown in Table
19.
TABLE-US-00021 TABLE 19 Bioavailability of SOF/Ledipasvir fixed
dose combination and single agent tablets Total Tablet SOF
Ledipasvir weight/ AUC.sub.inf C.sub.max AUC.sub.inf C.sub.max
Formulation Dose (mg) (ng*hr/mL) (ng/mL) (ng*hr/mL) (ng/mL) Signle
Agent SOF 400 mg 11900 Control SOF Ledipasvir 90 (23.5) 764 (27.3)
9620 (45.6) 314 (40.5) tablet + mg Ledipasvir SD tablet
SOF/Ledipasvir 12500 784 (36.2) 9570 (46.6) 314 (45.2) FDC Tablet
(23.1)
[0278] These results, therefore, show that SOF/ledipasvir fixed
dose combination (co-granulated) and single agent tablets are
bioequivalent.
Example 8. Solubility Studies for Amorphous Ledipasvir
[0279] This example examines the physicochemical properties
different ledipasvir forms, including amorphous and crystalline
free base, solvates, and salts, with respect to solubility.
A. Materials and Methods
[0280] pH-Solubility Profile
[0281] The aqueous solubility of ledipasvir amorphous free base was
determined across the pH range of 1 to 10. Excess solid ledipasvir
was added to a range of pH-adjusted aqueous solutions (titrated
with HCl or NaOH) and stirred for 48 hours at room temperature. The
suspensions were then filtered through regenerated cellulose
syringe filters. The pH value of the supernatant was measured, and
the supernatant was diluted as appropriate with 50:50 H.sub.2O+0.1%
TFA:ACN and assayed for ledipasvir content by the HPLC-UV
method.
Solubility in Simulated Intestinal Media
[0282] Solubility of ledipasvir amorphous free base was assessed in
three types of simulated intestinal fluids at pH 6.5 or pH 5.0; and
simulated intestinal bile salt and lecithin mixture (SIBLM), pH
6.4. Excess solid ledipasvir was added to the respective SIFs and
stirred for 48 hours at room temperature. The resulting suspensions
were then filtered through regenerated cellulose syringe filters.
The supernatant was diluted as appropriate with 50:50 H.sub.2O+0.1%
TFA:ACN and assayed for ledipasvir content by the HPLC-UV
method.
Excipient Solubility
[0283] Solubility of ledipasvir amorphous free base and ledipasvir
crystalline D-tartrate was measured in a wide range of
pharmaceutically acceptable solvents, including cosolvents,
surfactants, fatty acids, triglycerides, or blends thereof.
Material was weighed into scintillation vials and stirred for up to
48 hours at room temperature. In many cases, solubility was higher
than the amount of solid used in the sample, thus many results are
reported as `greater than` or `greater than or equal to` if the
concentration was not quantitatively determined by HPLC-UV.
[0284] Additionally, aqueous solubility was measured as a function
of time in the presence of 0.1% w/w surfactants and polymers in pH
2 (50 mM citrate) and pH 5 (50 mM citrate). ledipasvir crystalline
forms (acetone solvate Form II; anhydrous FB Form III; D-tartrate)
and amorphous form were evaluated to identify differences in
dissolution behavior. Excess solid was added to aqueous buffered
solutions; samples were withdrawn at predetermined intervals (2, 5,
8, 10, 15, 20, 30, 45, 60 minutes, and 24 hours), filtered through
regenerated cellulose filters, and diluted for concentration
measurement by the HPLC-UV method.
B. Results
Solubility and Dissolution Rate
[0285] The pH-solubility profiles of all available ledipasvir forms
were determined at room temperature and are graphically shown in
FIG. 9. The flat portion of the solubility profile (pH >5)
represents the intrinsic aqueous solubility of the free base. The
aqueous solubility of ledipasvir significantly increases as the pH
of the solution is lowered below the pKa of the ionizable groups.
All forms lose crystallinity, reverting to the amorphous free base
in aqueous solution, and thus show similar aqueous solubility
properties at steady-state. However, dissolution properties are
form dependent and are described in further detail below.
Ledipasvir Amorphous Free Base
[0286] The intrinsic solubility of ledipasvir amorphous free base
(FB) is approximately 0.04 .mu.g/mL. Under acidic conditions, the
solubility increases to 1 mg/mL at pH 2.3, and peaks at about 7
mg/mL at pH 1.6, as shown in Table 20 and FIG. 9. Solubility of
ledipasvir in simulated intestinal fluids is governed by both the
pH of the medium and the presence of bile salts and lecithin. In
fasted state simulated intestinal fluids (FaSSIF) at pH 6.5 and
room temperature, the solubility is 0.025 mg/mL, and this is
increased approximately 10-fold to 0.232 mg/mL in simulated bile
and lecithin mixture (SIBLM, pH 6.5) due to the increased
concentration of bile salts and lecithin. A similar solubility
enhancement to 0.230 mg/mL is observed in fed state simulated
intestinal fluid (FeSSIF, pH 5), containing lower bile salt and
lecithin mixtures than SIBLM. The solubility increase in this
mixture is predominantly attributed to the ionization state of the
molecule at pH 5.
TABLE-US-00022 TABLE 20 Solubility of Ledipasvir amorphous free
base as a function of pH at room temperature Aqueous Media
Solubility (mg/mL) Aqueous, pH 1.6 (HCl) 6.855 Aqueous, pH 2.3
(HCl) 1.096 Aqueous, pH 3.1 (HCl) 0.0132 Aqueous, pH 4.1 (HCl)
0.00011 Aqueous, pH 5.5 (HCl) 0.00003 Aqueous, pH 6.2 (unaltered)
0.00003 Aqueous, pH 7.2 (NaOH) 0.00001 FaSSIF.sup.1 pH = 6.5 0.025
FeSSIF.sup.2 pH = 5.0 0.230 SIBLM.sup.3 pH = 6.4 0.232 .sup.1FaSSIF
is water with 3 mM sodium taurocholate and 0.75 mM lecithin, pH
adjusted to 6.5 with phosphate buffer, ionic strength adjusted to
0.15M with NaCl. .sup.2FeSSIF is water with 15 mM sodium
taurocholate and 3.75 mM lecithin, pH adjusted to 6.5 with
phosphate buffer, ionic strength adjusted to 0.15M with NaCl.
.sup.3SIBLM is water with 30 mM sodium glycocholate, 30 mM sodium
glycochenodesoxycholate, 15 mM sodium glycodesoxycholate, 10 mM
sodium taurocholate, 10 mM sodium taurochenodesoxycholate, 5 mM
sodium taurodesoxycholate, 50 mM sodium chloride, and 11 mM
lecithin, pH adjusted to 6.4 with phosphate buffer, ionic strength
adjusted to 0.15M with NaCl.
[0287] The dissolution rate of ledipasvir amorphous free base at pH
3 and 6 was also tested. At pH 3, the dissolution of the amorphous
free base form is faster than that of the crystalline free base and
acetone solvate forms. However, at pH 6, all free base forms show
similar dissolution rate profiles.
[0288] As shown in Table 21, ledipasvir amorphous free base is
freely soluble (>500 mg/mL) in ethanol and other organic
solvents such as propylene glycol and PEG 400. Its solubility is
greater than 200 mg/mL in surfactants (e.g., polysorbate 80,
Cremophor EL, Labrasol) and lipid blends. Its solubility in oleic
and octanoic acids is greater than 500 mg/mL. Solubility of
ledipasvir in short-chain triglycerides (SCTs, tributyrin) is
limited to 20 mg/mL, and decreases to less than 1 mg/mL in
long-chain triglycerides (LCTs, soybean oil). It has a solubility
of 25 mg/mL in the vehicle chosen for toxicological studies: 45%
propylene glycol, 15% caprylocaproyl macrogol-8 glycerides (Solutol
HS 15.degree.), and 40% water (pH 2.5 by HCl).
TABLE-US-00023 TABLE 21 Solubility of Ledipasvir free base forms
and Ledipasvir D-tartrate in organic solvents and excipients at
room temperature Solubility (mg/mL) Crystalline Anhydrous
Crystalline Acetone Crystalline D-tartrate Amorphous Solvate Free
Base Salt Free Base (Ledipas- (Ledipasvir (Ledipas- (Ledipasvir)
vir-03) Form III) vir-02) Acetone 5 5 -- <1 Acetonitrile >500
-- -- 12 Methanol >500 >500 -- 23 95% Methanol + -- -- -- 19
5% water Ethanol >500 >500 >500 4 95% Ethanol + -- -- -- 5
5% water PEG 400 >500 >500 >500 4 Propylene glycol >500
>500 >500 6 Octanoic acid >500 >500 -- <1 Oleic acid
>500 >500 >500 <1 Polyoxyl 35 >200 -- -- Castor Oil
(Cremophor EL) Polysorbate 80 >200 >200 >200 3 (Tween 80)
Caprylocaproyl >300 >300 >300 3 macrogolglycerides
(Labrasol) Tributyrin 9 -- -- -- Soybean oil 2 2 2 -- RSSEDDS.sup.1
>500 -- -- 10 .sup.1RSSEDDS: 10% Ethanol, 10% PG, 40% Solutol
HS-15, 40% Labrasol
[0289] Dilute nonionic surfactants generally increase ledipasvir
solubility at both pH 2 and 5, as presented in Table 22. Similar
effects were observed with nonionic polymers, though to a lesser
extent. Sodium lauryl sulfate (SLS), an anionic surfactant,
improves the solubility of ledipasvir at pH 5. However, a
significant decrease in solubility is noted in presence of SLS
under acidic conditions (pH 2). This observation is consistent with
weakly basic compounds that have low intrinsic aqueous solubility,
presumably forming an insoluble estolate salt.
TABLE-US-00024 TABLE 22 Solubility of Ledipasvir amorphous free
base in surfactant or polymeric excipients (0.1% w/w) diluted into
aqueous media at pH 2 and 5 at room temperature Excipient (0.1% w/w
Solubility (mg/mL) in aqueous media) pH 2 pH 5 No excipient 4.94
0.0001 Sodium lauryl sulfate 0.05 0.243 Labrasol 7.44 -- Cremaphor
EL 9.11 0.0699 Polysorbate 80 9.27 0.0624 Poloxamer 188 7.19 0.0005
HPC (hydroxypropylcellulose) 4.67 0.0001 HPMC
(hydroxymethylcellulose) 5.27 0.0003 PVP (povidone) 5.47 0.0004
PVP/VA (copovidone) 6.73 0.0010
Ledipasvir Crystalline Acetone Solvate (Ledipasvir-03)
[0290] The ledipasvir acetone solvate (ledipasvir-03) showed
similar steady-state solubility as the other forms. Ledipasvir-03
has the slowest dissolution of all forms tested. Its dissolution at
pH 6 was indistinguishable from that of other forms due to poor
intrinsic solubility (<0.1 .mu.g/mL).
[0291] Ledipasvir-03 is soluble in many organic solvents and
pharmaceutically acceptable solvents, and the solubilities are
comparable to those listed for ledipasvir amorphous free base, as
also shown in Table 21.
Ledipasvir Crystalline Free Base (Form III)
[0292] Ledipasvir crystalline free base Form III showed similar
steady-state solubility as the other forms (FIG. 9). This form
dissolves more slowly than the amorphous free base, but faster than
ledipasvir-03. Dissolution at pH 6 was indistinguishable from that
of the other forms due to poor intrinsic solubility (<0.1
.mu.g/mL). Solubility in a wider range of organic vehicles has not
been explored, though is anticipated to be similar to other free
base forms.
Ledipasvir Crystalline D-Tartrate Salt (Ledipasvir-02)
[0293] Ledipasvir crystalline D-tartrate salt (ledipasvir-02)
showed similar steady-state solubility as the other forms (FIG. 9).
Dissolution behavior ledipasvir-02 is improved relative to all free
base forms. At pH 3, ledipasvir-02 shows a roughly 5- to 10-fold
faster initial dissolution rate than the free base forms, and
roughly doubled the amount of ledipasvir in solution through 60
minutes compared to the amorphous form. At pH 6, the increased
dissolution rate was also apparent. However, rapid dissociation of
the salt at this pH resulted in equivalent solubility values to
other forms within minutes.
[0294] Ledipasvir-02 is not soluble in various organic media, as
shown in Table 21. Maximal solubility of ledipasvir-02 in any
organic vehicle is 20 mg/mL in methanol; this limits the use of
ledipasvir-02 in solubilized formulations or processes that require
solubilization in organic media.
[0295] Ledipasvir has low aqueous solubility and high permeability,
and is considered a BCS Class 2 compound. The data presented in
this example indicate that in water, all forms of ledipasvir: the
amorphous free base, crystalline free base acetone solvate
(ledipasvir-03), crystalline anhydrous free base (Form III), and
crystalline D-tartrate salt (ledipasvir-02), convert to the
amorphous free base, and have similar aqueous solubility at steady
state. The aqueous solubility of ledipasvir is less than 0.1
.mu.g/mL in its neutral form (pH >5), but substantially
increases under acidic conditions due to protonation of two basic
moieties. The aqueous dissolution rate of ledipasvir amorphous free
base is faster than that of crystalline free base forms. However,
all free base forms have slower dissolution rates than the
crystalline D-tartrate salt (ledipasvir-02). Ledipasvir-02 also
shows improved wetting in aqueous media. ledipasvir free base
forms, crystalline and amorphous, are highly soluble in a range of
cosolvents and surfactants. In contrast, ledipasvir-02 is poorly
soluble in organic excipients, and this property potentially limits
its utility.
[0296] Ledipasvir amorphous free base was used in Phase 1 clinical
studies, but drug substance manufacturing was identified as a
critical limitation of the form. Ledipasvir crystalline D-tartrate
salt (ledipasvir-02) was then identified as part of a more
extensive salt and form screen and was used in Phase 2, however,
poor solubility in organic excipients limits its utility in
non-conventional formulations. Crystalline ledipasvir acetone
solvate (ledipasvir-03) is used to develop a spray dried dispersion
formulation to support future clinical studies due to its
solubility in organic solvents and excipients relative to
crystalline ledipasvir D-tartrate salt and improved
manufacturability over the other free base forms.
Example 9: Efficacy of a Fixed Dose Combination of Sofosbuvir and
Ledipasvir with and without Ribavirin in Patients with HCV
Infections
[0297] Patients with HCV infections were treated with the fixed
dose combination of sofosbuvir and ledipasvir, with and without
ribavirin. Patients used in the studies include those that were
treatment naive (non-cirrhotic), i.e. had not previously been
treated for HCV, and those that were prior protease-inhibitor (PI)
failures and null responders (with and without cirrhosis), i.e. had
previously been treated for HCV but failed to respond to the
treatment. The treatment naive pateints were treated for 6, 8, and
12 weeks and the null responders were treated for 12 weeks.
Study 1
[0298] Cohort 1 of study 1 included treatment-naive, Genotype-1
patients without cirrhosis. The patients were randomized 1:1:1 into
three groupds to receive 1) SOF/ledipasvir fixed dose combination
for 8 weeks, 2) SOF/ledipasvir fixed dose combination with
ribavirin for 8 weeks, or 3) SOF/ledipasvir fixed dose combination
for 12 weeks (FIG. 10).
[0299] Cohort 2 of study 1 included Protease-Inhibitor
treatment-experienced, Genotype-1 patients (Prior
Protease-Inhibitor treatment failures, 50% of whom had compensated
cirrhosis). The pateints were randomized to receive 12 weeks of: 1)
SOF/ledipasvir fixed dose combination or 2) SOF/ledipasvir fixed
dose combination with ribavirin (FIG. 10). In Cohort 2, the
patients must not have discontinued prior therapy due to an adverse
event.
[0300] In study 1, there was a broad inclusion criteria, namely,
there was no upper limit to age or BMI. Platelets were
>50,000/mm.sup.3. The demographics of study 1 are shown in Table
23, below.
TABLE-US-00025 TABLE 23 Demographics SOF/Ledipasvir fixed dose
combination .+-. ribavirin (Cohort 1 and 2) N = 100 Mean age, y
(range) 50 (21-73) Male, n (%) 66 (66) Black, n (%) 9 (9) Hispanic,
n (%) 40 (40) Mean BMI, kg/m.sup.2 (range) 29.9 (18-48) IL28B CC, n
(%) 15 (15) GT 1a, n (%) 87 (87) Mean baseline HCV RNA, 6.1
(3.7-7.2) log.sub.10 IU/mL (range) Cohort 2 (N = 40) Cirrhosis, n
(%) 22/40 (55) Mean Platelet Count (.times.10.sup.3/.mu.L) 107 Mean
Albumin (g/dL) 3.8
[0301] Of 100 patients enrolled in study 1, 97% achieved sustained
viral response. Of the failures, two patients relapsed (one from
Group 1 (i.e., SOF/Ledipasvir x 8 Weeks) and one from Group 4
(i.e., SOF/ledipasvir.times.12 Weeks), and one patient was lost to
follow up from Group 3 (i.e., SOF/ledipasvir.times.12 Weeks).
However, the patient lost to follow up had achieved SVR at week 8
and declined further return visits.
[0302] In Cohort 1 of study 1 (i.e. Treatment Naive, Non-Cirrhotic
Patients), 58 out of the 60 patients treated for 8 or 12 Weeks
achieved SVR. In corhort 2 of study 1 (i.e. Treatment Experienced,
PI failure Patients), 39 out of the 40 patients treated for 12
Weeks achieved SVR12. 21 out of the 21 patients with cirrhosis
achieved SVR12 (FIG. 11).
[0303] In study 1, seven out of the nine patients with NS5A
Resistance Associated Variants (RAVs) achieved sustained viral
response. In addition, all patients with NS3/4A Resistance
Associated Variants achieved sustained viral response.
Interestingly, S282T mutation and multiple NS5A RAVs were detected
at relapse in the patient who failed from the Group 1 (Table 24).
The safety summary and a breakdown of the adverse effects are shown
in Tables 25 and 26, respectively.
TABLE-US-00026 TABLE 24 Resistance analysis SOF/Ledipasvir fixed
dose combination .+-. ribavirin NS5A RAVs, n % 9/100 (9) NS3/4A
RAVs, n % 29/40 (73)* *number of patients in Cohort 2 with prior
exposure to a protease inhibitor
TABLE-US-00027 TABLE 25 Safety Summary SOF/Ledipasvir
SOF/Ledipasvir fixed dose fixed dose combination combination +
Patients, n (%) N = 58 ribavirin N = 42 Overall AEs 24 (41%) 24
(57%) safety Grade 3-4 AEs 0 6 (14%) Serious AEs 2* (3%) 2** (5%)
Treatment 0 0 discontinuation due to AEs Laboratory Grade 3-4
laboratory 4 (7%) 6 (14%) abnormalities abnormality Hemoglobin 0 8
(19%) <10 g/dL Hemoglobin 0 2 (5%) <8.5 g/dL *peptic ulcer,
spinal compression fracture **delerium, suicidal ideation
TABLE-US-00028 TABLE 26 Adverse Events ( .gtoreq.5% of patients
overall) SOF/Ledipasvir fixed dose SOF/Ledipasvir fixed dose
Preferred term, combination combination + ribavirin n (%) N = 58 N
= 42 Any adverse 24 (41%) 24 (57%) event Nausea 3 (5%) 6 (14%)
Anemia 0 8 (19%) Upper Resp 4 (7%) 4 (10%) Tract Infx Headache 3
(5%) 4 (10%)
Study 2
[0304] In study 2, the treatment-naive patients received
SOF/ledipasvir fixed dose combination with ribavirin and prior null
responders, all of whom had cirrhosis, were randomized to receive
twelve weeks of: 1) SOF/ledipasvir fixed dose combination or 2)
SOF/ledipasvir fixed dose combination with ribavirin.
Results
[0305] Of the 144 patients treated in both studies 1 and 2, 136 out
of 144 (94%) achieved SVR at four weeks post treatment. Of the 85
treatment-naive patients in these two studies, three of 25 patients
failed to achieve SVR after 6 weeks of SOF/ledipasvir fixed dose
combination with ribavirin therapy, whereas 100% (60/60) patients
achieved SVR after 8 or 12 weeks of SOF/ledipasvir fixed dose
combination with and without ribavirin therapy. Of the 59
treatment-experienced patients in these two studies, three
cirrhotic patients relapsed after receiving 12 weeks of
SOF/ledipasvir fixed dose combination without ribavirin.
Conversely, no virologic failures were observed in the
SOF/ledipasvir fixed dose combination with ribavirin treatment
groups, but two patients in these groups were lost to follow-up.
SOF/ledipasvir fixed dose combination with and without ribavirin
was well tolerated, with few SAEs and minimal adverse events.
Conclusion
[0306] SOF/ledipasvir fixed dose combination +/- ribavirin may be
given for as little as 8 weeks to treatment-naive non-cirrhotic
patients. Treatment-experienced patients, even those with
cirrhosis, achieved high SVR rates with 12 weeks of the of
SOF/ledipasvir fixed dose combination with and without ribavirin
therapy.
Example 10: Efficacy of Multiple Anti-HCV Combination Therapy in
Chronically Infected Hepatitis C Patients
[0307] To evaluate the safety, tolerability, and efficacy of 4 to
12 weeks of SOF with ledipasvir, alone or in combination with
Compound E and/or Compound J in patients with HCV, patients with
HCV will be dosed as shown in Table 27.
TABLE-US-00029 TABLE 27 Dosing Group Treatment Dosing Patient
description Group A 12 weeks of SOF with ledipasvir (400 Patients
(n = 20) SOF/ledipasvir mg/90 mg respectively once a monoinfected
with day in a fixed dose HCV genotype 1 combination) administered
who are HCV orally for 12 weeks treatment naive Group B 6 weeks of
SOF with ledipasvir (400 Patients (n = 20) SOF/ledipasvir/ mg/90 mg
respectively once a monoinfected with Compound E day in a fixed
dose HCV genotype 1 combination) in combination who are HCV with
Compound E (500 mg treatment naive once daily) for 6 weeks Group C
6 weeks of SOF with ledipasvir (400 Patients (n = 20)
SOF/ledipasvir/ mg/90 mg respectively once a monoinfected with
Compound J day in a fixed dose HCV genotype 1 combination) in
combination who are HCV with Compound J (80 mg once treatment naive
daily) for 6 weeks. Group D 12 weeks of SOF with ledipasvir (400
Patients (n = up to 25) SOF/ledipasvir mg/90 mg respectively once a
monoinfected with day in a fixed dose HCV genotype 1 combination)
administered who were previously orally for 12 weeks treated in
Group B, C, F, G or H of this study or a similar study Group E 12
weeks of SOF with ledipasvir (400 Patients (n = 20) SOF/ledipasvir
mg/90 mg respectively once a monoinfected with day in a fixed dose
HCV genotype 4 combination) administered who are HCV orally for 12
weeks treatment naive or treatment experienced Group F 6 weeks of
SOF with ledipasvir (400 Patients (n = 50) SOF/ledipasvir/ mg/90 mg
respectively once a monoinfected with Compound J day in a fixed
dose HCV genotype 1 combination) in combination with advanced liver
with Compound J (80 mg once disease who are daily) for 6 weeks. HCV
treatment naive (n = 25) or treatment experienced (n = 25) Group G
4 weeks of SOF with ledipasvir (400 Patients (n = 25)
SOF/ledipasvir/ mg/90 mg respectively once a monoinfected with
Compound J day in a fixed dose HCV genotype 1 combination) in
combination who are HCV with Compound J (80 mg once treatment
naive, daily) for 4 weeks Stage 0-2 liver disease Group H 4 weeks
of SOF with ledipasvir (400 Patients (n = 25) SOF/ledipasvir/ mg/90
mg respectively once a monoinfected with Compound J/ day in a fixed
dose HCV genotype 1 Compound E combination) in combination who are
HCV with Compound J (80 mg once treatment naive, daily) and
Compound E (250 Stage 0-2 liver mg once daily) for 4 weeks
disease
[0308] The primary analysis set for safety analyses will include
patients who received at least one dose of study drug. On treatment
data will be analyzed and defined as data collected from the first
dose of study drug through the date of last dose of study drug plus
30 days. Patients who receive study drug other than that to which
they were assigned will be analyzed according to the study drug
received.
[0309] The analysis set for antiviral activity analyses will
include patients who were enrolled into the study and received at
least one dose of study drug.
[0310] The pharmacokinetic analysis set will include all patients
who are enrolled and have received at least one dose of study
medication.
[0311] The patient will be started on study treatment after
confirming eligibility on Day 0 and after being informed fully
about the remainder of the study, and then signing the specific
consent for the treatment group (if not done previously). Blood
will be drawn for HCV viral loads, study drug levels, lipid levels
for research if not already drawn during screening, immunologic
studies, and for storage prior to dosing as part of the screening
consent. A pregnancy test will be done for females with
childbearing potential and the pregnancy test must be negative on
Day 0 prior to dosing with study drugs. Patients may be asked to
fill out a baseline adherence questionnaire and an electronic pill
bottle cap, which records pill bottle openings will be placed on
all study drug bottles. Assistance will be provided filling out the
questionnaire as needed. Patients on Arms B and H, will also be
provided with a diary at Day 0, Week 2, Week 4 (Arm B only) on
which to record gastrointestinal side effects.
[0312] On arrival at the clinic for scheduled study visits,
patients will have their vital signs obtained, females will undergo
a pregnancy test (if appropriate per schedule and of childbearing
potential), clinical laboratories drawn, and a review of the study
restrictions.
[0313] At each scheduled study visit (does not include Day 1, 3, 5,
10, Week 2, Week 3, Week 6 (not applicable for arm F, G or H) or
post-treatment week 2 and 8 which are for lab collection only),
patients will be asked about their state of health and use of any
concomitant medication since the previous study visit. They will
also be questioned about adverse events and their adherence with
study restrictions. Vital signs, weight and examination will be
performed as per the study flow. A complete list of study
procedures and lab tests to be performed is in the Schedule of
Tests, below. In addition, patients may be seen at unscheduled
visits for a grade 3 or 4 adverse event or any unexpected adverse
event or potential toxicity.
[0314] Patients may be asked to fill out a follow-up adherence
questionnaire and pill bottle openings may be recorded from the
electronic bottle cap at Day 7 (Group A), Week 4 (Group A), Week 6
(Groups B and C), Week 8 (Group A), and Week 12 (Group A).
Assistance will be provided filling out the questionnaire as
needed.
[0315] Patients in Groups B and H will be asked to bring their
side-effect diaries to visits on Week 2, 4, 6 (B only).
[0316] Some of the visits have a small amount of flexibility
regarding when they need to occur. Visits occurring during the
interval when the patient is receiving study drug have limited
flexibility since they occur so frequently, so a visit skipped
during this period may be considered a missed visit. The window
period for visit schedules is as shown in Table 28.
TABLE-US-00030 TABLE 28 Window Period for Visit Schedule For 12
week regimen, Group A: Days 0, 1, 3 Days 5, 7, 10, Weeks 3, 4, 6
Weeks 8, 12 (no window) 14 (+/-2 days) (+/-3 days) (+/-5 days)
Optional Week 12 Research Liver Biopsy (+/-14 days) For 6 week
regimens, Groups B & C: Days 0, 1, 3 Days 5, 7, 10, Weeks 3, 4,
6 (no window) 14 (+/-2 days) (+/-3 days) Optional Week 6 Research
Liver Biopsy (+/-14 days) For 12 week regimens, Groups D and E: Day
0 Week 4 Weeks 8, 12 (no window) (+/-3 days) (+/-7 days) For 6 week
regimen, Group F: Day 0 Weeks 2, 4 Week 6 (+/-5 days) (no window)
(+/-3 days Optional Week 6 Research Liver Biopsy (+/-14 days) For 4
week regimen, Group G or H: Day 0 Day 7 Weeks 2, 4 (+/-3 days) (no
window) (+/-2 days) Optional Week 4 Group H only Research Liver
Biopsy (+/-14 days)
[0317] During the four week visit, HCV RNA may be obtained to
determine if virologic-response based treatment stopping criteria
have been met. Patients who fail to achieve >2 log 10 HCV RNA
drop at this time (unless >2 log drop would be below LLOQ)
should be discontinued from therapy unless a review by the
PI/LAI/Sponsor Medical Monitor determines otherwise (see
9.3.1).
[0318] At the end of treatment duration as determined by the study
group, patients may discontinue dosing of SOF and ledipasvir,
Compound E, and/or Compound J. In addition, if a patient's
participation terminates prior to completion of pre-specified study
drug duration, the End of Treatment assessments may be performed at
any end-of-treatment visit. An optional research liver biopsy for
research purposes may be performed at this time in up to 10
patients in each study group. The additional liver biopsy data will
serve to explore hepatic HCV RNA sequence analysis. If patients are
undergoing the optional research liver biopsy, they may have safety
labs completed prior to the procedure and imaging as medically
indicated. Patients who have a HCV VL<LLOQ may receive education
about how to prevent re-infection with HCV.
[0319] All patients may be assessed for sustained virologic
response at the 12 Weeks Post End of Treatment visit. Patients who
have HCV VL<LLOD may be provided with education about how to
prevent re-infection with HCV.
[0320] After discontinuation of the study drug, patients may be
followed at 2, 4, 8, 12, 24, 36, and 48 weeks post-end of
treatment. A serum pregnancy test may be done with each visit, as
appropriate. Week 2 and 8 Post-End of Treatment may include only
collection of labs.
[0321] Subjects (n=18) received single doses of sofosbuvir (400 mg)
alone or in combination with Compound E (500 mg QD) under fed
conditions. Preliminary PK results for the combination of
sofosbuvir with Compound E are presented in Table 29 and
demonstrate lack of a clinically significant interaction between
sofosbuvir and Compound E.
TABLE-US-00031 TABLE 29 Pharmacokinetic Data for SOF, Compound E,
and ledipasvir alone and upon co-administration SOF (n = 18) Mean
(% CV) SOF alone SOF + Compound E % GMR (90% CI) AUCinf 921 (61.2)
1150 (40.2) 135 (116, 159) (ng.hr/ml) SOF + Compound E + ledipasvir
% GMR (90% CI) 2560 (42.9) 297 (253, 348) SOF + Compound E % GMR
(90% CI) AUClast 908 (62.2) 1140 (41.4) 135 (115, 159) (ng.hr/ml)
SOF + Compound E + ledipasvir % GMR (90% CI) 2550 (43.1) 301 (255,
354) SOF + Compound E % GMR (90% CI) Cmax (ng/ml) 515 (78.3) 587
(51.1) 130 (97.1, 175) SOF + Compound E + ledipasvir % GMR (90% CI)
1260 (55.1) 283 (219, 366)
[0322] It should be understood that although the present invention
has been specifically disclosed by preferred embodiments and
optional features, modification, improvement and variation of the
inventions embodied therein herein disclosed may be resorted to by
those skilled in the art, and that such modifications, improvements
and variations are considered to be within the scope of this
invention. The materials, methods, and examples provided here are
representative of preferred embodiments, are exemplary, and are not
intended as limitations on the scope of the invention.
[0323] The invention has been described broadly and generically
herein. Each of the narrower species and subgeneric groupings
falling within the generic disclosure also form part of the
invention. This includes the generic description of the invention
with a proviso or negative limitation removing any subject matter
from the genus, regardless of whether or not the excised material
is specifically recited herein.
[0324] In addition, where features or aspects of the invention are
described in terms of Markush groups, those skilled in the art will
recognize that the invention is also thereby described in terms of
any individual member or subgroup of members of the Markush
group.
[0325] All publications, patent applications, patents, and other
references mentioned herein are expressly incorporated by reference
in their entirety, to the same extent as if each were incorporated
by reference individually. In case of conflict, the present
specification, including definitions, will control.
* * * * *