U.S. patent application number 12/855458 was filed with the patent office on 2011-04-21 for method of drug delivery for bone anabolic protein.
This patent application is currently assigned to Radius Health, Inc.. Invention is credited to Michael J. Dey, Bart Henderson, C. Richard Lyttle, Nathalie Mondoly, Benedicte Rigaud.
Application Number | 20110092425 12/855458 |
Document ID | / |
Family ID | 40873336 |
Filed Date | 2011-04-21 |
United States Patent
Application |
20110092425 |
Kind Code |
A1 |
Dey; Michael J. ; et
al. |
April 21, 2011 |
METHOD OF DRUG DELIVERY FOR BONE ANABOLIC PROTEIN
Abstract
The present invention provides a storage-stable composition
containing a parathyroid hormone-related protein (PTHrP) and
methods of using a PTHrP and the PTHrP compositions described
herein to treat osteoporosis, to increase bone mass or to increase
bone quality. The composition is storage stable, in sterile form,
and in general may be stored at room temperature for at least
several weeks to allow convenient parenteral administration to
human patients.
Inventors: |
Dey; Michael J.; (Sanbach,
GB) ; Mondoly; Nathalie; (Le Chesnay, FR) ;
Rigaud; Benedicte; (Oulins, FR) ; Henderson;
Bart; (Belmont, MA) ; Lyttle; C. Richard;
(Bala Cynwyd, PA) |
Assignee: |
Radius Health, Inc.
Cambridge
MA
Ipsen Pharma S.A.S
Boulogne-Billancourt
|
Family ID: |
40873336 |
Appl. No.: |
12/855458 |
Filed: |
August 12, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12151975 |
May 9, 2008 |
7803770 |
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12855458 |
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PCT/US07/21216 |
Oct 3, 2007 |
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12151975 |
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60848960 |
Oct 3, 2006 |
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Current U.S.
Class: |
514/11.8 |
Current CPC
Class: |
A61K 38/29 20130101;
A61K 9/0019 20130101; A61P 19/10 20180101; A61K 9/08 20130101 |
Class at
Publication: |
514/11.8 |
International
Class: |
A61K 38/29 20060101
A61K038/29; A61P 19/10 20060101 A61P019/10 |
Claims
1-48. (canceled)
49. A single or multi-dose sealed container, vial or cartridge that
contains an aqueous solution comprising at least 80 .mu.g of
[Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO: 2) wherein said
aqueous solution comprises an effective amount of buffer to
maintain the pH between 2 and 7.
50. The container, vial or cartridge of claim 49 wherein said
buffer is selected from the group consisting of acetate, tartrate,
phosphate and citrate buffers.
51. The container, vial or cartridge of claim 50 wherein said
buffer is an acetate buffer.
52. The container, vial or cartridge of claim 51 wherein said
buffer is acetic acid and sodium acetate.
53. The container, vial or cartridge of claim 49 wherein said pH is
maintained between 3 and 6.
54. The container, vial or cartridge of claim 49 wherein said pH is
maintained between 4 and 6.
55. The container, vial or cartridge of claim 49 wherein said pH is
maintained between 4.5 and 5.6.
56. The container, vial or cartridge of claim 55 further comprising
phenol.
57. The container, vial or cartridge of claim 56 wherein said
phenol is present in a concentration from about 0.25 to about 5
mg/mL.
58. A multi-dose sealed container, vial or cartridge that contains
an aqueous solution comprising at least 160 .mu.g of
[Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO: 2) wherein said
aqueous solution comprises an effective amount of buffer to
maintain the pH between 2 and 7.
59. The container, vial or cartridge of claim 58 wherein said
buffer is selected from the group consisting of acetate, tartrate,
phosphate and citrate buffers.
60. The container, vial or cartridge of claim 59 wherein said
buffer is an acetate buffer.
61. The container, vial or cartridge of claim 60 wherein said
buffer is acetic acid and sodium acetate.
62. The container, vial or cartridge of claim 58 wherein said pH is
maintained between 3 and 6.
63. The container, vial or cartridge of claim 58 wherein said pH is
maintained between 4 and 6.
64. The container, vial or cartridge of claim 58 wherein said pH is
maintained between 4.5 and 5.6.
65. The container, vial or cartridge of claim 64 further comprising
phenol.
66. The container, vial or cartridge of claim 65 wherein said
phenol is present in a concentration from about 0.25 to about 5
mg/mL
67. A multi-dose sealed container, vial or cartridge that contains
an aqueous solution comprising at least 160 .mu.g of
[Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO: 2) wherein said
aqueous solution comprises an effective amount of a buffer to
maintain the pH at about 5.1 and wherein said buffer is an acetate
buffer and wherein said aqueous solution further comprises about 5
mg/mL of phenol.
68. The container, vial or cartridge of claim 67 wherein said
[Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 is present in a concentration of
about 2 mg/mL.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 12/151,975, filed on May 9, 2008, which is a
continuation-in-part of International Application No.
PCT/US2007/021216, which designated the United States and was filed
on Oct. 3, 2007, published in English, which claims the benefit of
U.S. Provisional Application No. 60/848,960, filed on Oct. 3, 2006.
The entire teachings of the above applications are incorporated
herein by reference.
BACKGROUND OF THE INVENTION
[0002] Parathyroid hormone-related protein ("PTHrP") is a 139 to
173 amino acid-protein. PTHrP and certain analogs are known to be
useful to improve bone mass and quality in the treatment of
osteoporosis and related disorders. However, the commercial use of
these proteins as pharmaceutical agents requires the development of
a formulation that is acceptable in terms of storage stability and
ease of preparation.
[0003] Furthermore, currently available osteoporosis drugs have
limitations on suitable dosage ranges due to the unwanted
side-effects, such as hypercalcemia and increased stimulation of
bone resorption. These unwanted side-effects and resulting dose
limitations reduce the beneficial effects which can be achieved
from these drugs. Thus a need exists for compounds which can be
administered at a dose which will increase the beneficial effects
without an increase in the unwanted side-effects.
SUMMARY OF THE INVENTION
[0004] The present invention provides a storage-stable composition
containing a parathyroid hormone-related protein (PTHrP) and
methods of using PTHrPs and the PTHrP compositions described herein
to treat osteoporosis, to increase bone mass or to increase bone
quality. The composition is storage stable, in sterile form, and in
general may be stored at room temperature for at least several
weeks to allow convenient parenteral administration to human
patients.
[0005] In one embodiment, the present invention provides a
storage-stable composition suitable for administration to a subject
(e.g., a human). The composition comprises a PTHrP and an effective
amount of buffer to maintain the pH of the composition between 2
and 7. In a particular embodiment, the PTHrP is [Glu22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2).
[0006] In another embodiment, the present invention provides a
sealed container containing a storage-stable composition suitable
for administration to a subject. The composition comprises PTHrP or
an analog thereof and an effective amount of buffer to maintain the
pH of the composition between 2 and 7. In a particular embodiment,
the PTHrP is [Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2).
[0007] In another embodiment, the present invention provides a drug
delivery device comprising one or more than one single-use
container which comprises a storage stable composition comprising
PTHrP or an analog thereof and an effective amount of buffer to
maintain the pH of the composition between 2 and 7. In a particular
embodiment, the PTHrP is [Glu.sup.22,25, Leu.sup.23,28,31,
Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.:2).
[0008] In another embodiment, the present invention provides a drug
delivery device comprising one or more than one multi-use
container, which comprises a storage stable composition comprising
PTHrP or an analog thereof and an effective amount of buffer to
maintain the pH of the composition between 2 and 7. In a particular
embodiment, the PTHrP is [Glu.sup.22,25, Leu.sup.23,28,31,
Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2).
[0009] In another embodiment the present invention provides a
method of treating osteoporosis in a subject in need thereof
comprising administering to the subject [Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2) in an amount between 40 and 160 .mu.g.
[0010] In another embodiment the present invention provides a
method of increasing bone mass or increasing bone quality in a
subject in need thereof comprising administering to the subject
[Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) in an amount
between 40 and 160 .mu.g.
[0011] The PTHrP compositions of the invention exhibit storage
stability in terms of hormone composition and activity. These
compositions eliminate the need for chemical stabilizers and other
stabilization techniques, such as, lyophilization. Furthermore,
these compositions can be administered, in general, in higher
dosages than currently available osteoporosis drugs, with the
reduction or elimination of unwanted side-effects, such as,
hypercalcemia or stimulation of bone resorption. This has the
advantage of an increase in beneficial physiological effects due to
the increased dosages and can result in a reduction in the length
of treatment time.
BRIEF DESCRIPTION OF THE DRAWINGS
[0012] FIG. 1 is a graph showing the stability of SEQ ID NO. 2 over
24 months at 5.degree. C. and 25.degree. C. without any chemical
stabilizer.
[0013] FIG. 2 is a graph showing the stability of lyophilized SEQ
ID NO. 2 over 24 months at 5.degree. C. 25.degree. C. and
40.degree. C.
[0014] FIG. 3 is a graph showing the plasma levels of the bone
formation marker Procollagen type 1 N-propeptide (P1NP) (ng/mL)
through two days pre-dosing, seven days of dosing and seven days
post-dosing. A=5 .mu.g SEQ ID NO. 2; B=20 .mu.g SEQ ID NO. 2; C=40
.mu.g SEQ ID NO.2; D=80 .mu.g SEQ ID NO.2; P=Placebo.
[0015] FIG. 4 is a graph showing the plasma levels of the bone
resorption marker Serum C-telopeptide type-1 collagen (Ctx) (ng/mL)
through two days pre-dosing, seven days dosing and seven days
post-dosing. A=5 .mu.g SEQ ID NO. 2; B=20 .mu.g SEQ ID NO. 2; C=40
.mu.g SEQ ID NO.2; D=80 .mu.g SEQ ID NO.2; P=Placebo.
[0016] FIG. 5 is a graph showing the plasma levels of serum ionized
calcium (mmol/L) thro.mu.gh 24 hours post-first dose and 24 hours
post-seventh dose. A=5 .mu.g SEQ ID NO. 2; B=20 .mu.g SEQ ID NO. 2;
C=40 .mu.g SEQ ID NO.2; D=80 .mu.g SEQ ID NO.2; P=Placebo.
[0017] FIG. 6 is a graph showing the plasma levels of PTH (pG/mL)
through 24 hours post-first dose and 24 hours post-seventh day
dose. A=5 .mu.g SEQ ID NO. 2; B=20 .mu.g SEQ ID NO. 2; C=40 .mu.g
SEQ ID NO.2; D=80 .mu.g SEQ ID NO.2; P=Placebo.
DETAILED DESCRIPTION OF THE INVENTION
[0018] As used herein "PTHrP" includes analogs and fragments of
native human PTHrP. An analog of PTHrP refers to a polypeptide
having between about 1 and about 20, between about 1 and about 15,
or between about 1 and about 10 art-accepted substitutions,
additions or insertions relative to human parathyroid
related-hormone protein (hPTHrP), or combinations thereof, not to
exceed a total combination of 20 substitutions, additions and
insertions. As used herein insertions, include the insertion of an
amino acid between two existing amino acids in the peptide chain.
As used herein addition means the addition of an amino acid to the
N or C terminus of the peptide chain. As used herein substitution
means the substitution of an amino acid for an existing amino acid
in the peptide chain. As used herein, "art-accepted" substitutions,
insertions or additions are those which would maintain or increase
and the biological and/or hormonal activity of the peptide and
which would not adversely affect the biologically activity of the
peptide. Art-accepted includes, for example, substitution of one
amino acid with a chemically or biologically similar amino acid,
such as a substituting one hydrophobic amino acid for another
hydrophobic amino acid. The PTHrPs are described with reference to
their variation from the native sequence of human parathyroid
hormone-related protein (hPTHrP).
[0019] A fragment of PTHrP refers to a polypeptide having a
sequence comprising less than the full complement of amino acids
found in PTHrP which, however, elicits a similar biological
response. The truncated PTHrP fragments may also be analogs as
defined above and need not be fully homologous with native PTHrP to
elicit a similar biological response.
[0020] Typically, the truncated analogs or fragments for use in the
methods and compositions of the present invention will be truncated
from the C-terminus and will have range from 30 to 40 residues. In
particular, hPTHrP(1-34) and analogs with between 1 and 15
substitutions thereof are useful in the methods and compositions of
the present invention.
[0021] The sequence of native hPTHrP (1-34) is as follows:
TABLE-US-00001 (SEQ ID NO: 1) Ala Val Ser Glu His Gln Leu Leu His
Asp Lys Gly Lys Ser Ile Gln Asp Leu Arg Arg Arg Phe Phe Leu His His
Leu Ile Ala Glu Ile His Thr Ala.
[0022] In a particular embodiment, the PTHrP is [Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2).
[0023] Other PTHrPs are described in U.S. Pat. No. 6,921,750,
5,955,574, 6,544,949, 5,723,577, and 5,696,095 the entire contents
of each of which are incorporated herein by reference.
[0024] A "buffer" as used herein is any acid or salt combination
which is pharmaceutically acceptable and capable of maintaining the
composition of the present invention within a desired pH range.
Buffers in the disclosed compositions maintain the pH in a range of
about 2 to about 7, about 3 to about 6, about 4 to about 6, about
4.5 to about 5.6, or about 5.1. Suitable buffers include, any
pharmaceutical acceptable buffer capable of maintaining the above
pH ranges, such as, for example, acetate, tartrate phosphate or
citrate buffers. In one embodiment, the buffer is an acetate or
tartrate buffer. In another embodiment the buffer is an acetate
buffer. In one embodiment the buffer is acetic acid and sodium
acetate.
[0025] In the disclosed compositions the concentration of buffer is
typically in the range of about 0.1 mM to about 1000 mM, about 0.2
mM to about 200 mM, about 0.5 mM to about 50 mM, about 1 mM to
about 10 mM or about 6 mM.
[0026] As used herein, an anti-microbial agent is a
pharmaceutically acceptable preservative, suitable for
administration to a subject, which inhibits, prevents or delays the
growth or micro organisms including, for example bacteria, viruses
and fungi in the compositions of the present invention. Suitable
anti-microbial agents for use in the compositions and methods of
the present invention include, but are not limited to, cresols,
benzyl alcohol, phenol, benzalkonium chloride, benzethonium
chloride, chlorobutanol, phenylethyl alcohol, methyl paraben,
propyl paraben, thiomersal and phenylmercuric nitrate and acetate.
In one embodiment the anti-microbial agents is m-cresol,
chlorocresol or phenol. In another embodiment the anti-microbial
agents is chlorocresol or phenol. In another embodiment the
anti-microbial agents is phenol.
[0027] As used herein an effective amount of an anti-microbial
agent is an amount effective to inhibits, prevents or delays the
growth or micro organisms including, for example bacteria, viruses
and fungi in the compositions of the present invention. In the
compositions of the present invention, the amount of anti-microbial
agent is typically in the range from about 0.1 to about 20 mg/ml,
about 0.2 to about 30 mg/ml, about 0.2 to about 10 mg/ml, about
0.25 to about 5 mg/ml, about 0.5 to about 50 mg/ml, about 1 to
about 10 mg/ml, about 3 mg/ml or about 5 mg/ml.
[0028] The compositions of the present invention typically are
ready to administer, aqueous solutions which are sterile,
storage-stable and pharmaceutically acceptable without the need for
reconstitution prior to administration. The compositions of the
present invention are suitable for administration to a subject
which means that they are pharmaceutically acceptable, non-toxic,
do not contain any components which would adversely affect the
biological or hormonal effects of the peptide. The compositions of
the present invention do not, for example, comprise any cells.
[0029] As used herein a composition of the present invention is
storage-stable if the amount, purity of the PTHrP remains above
about 95% of the original amount under one of the following
conditions: (1) storage for over 2 years at 5.degree. C.; or (2)
storage for over 30 days at 25.degree. C.
[0030] The compositions are typically stored in a sealed container,
vial or cartridge which is typically suitable for long term
storage. "Suitable for long-term storage" means that the vial,
container or cartridge does not allow for the escape of components
of the compositions of the present invention or the ingress of
external components, such as, micro organisms when kept for at
least 3 months at 25.degree. C.
[0031] The compositions of the present invention can be
administered by injection, typically subcutaneous injection.
[0032] The compositions of the present invention, can be stored in
single-dose or multi-dose sealed containers, vials or cartridges.
The sealed container, vial or cartridge is typically suitable for
use with a single or multi-dose injection pen or drug delivery
device, which typically allows the patient to administer the
peptide themselves. The sealed container can comprise one or more
doses of the peptide of the present invention, wherein each dose
comprises an effective amount of the peptide as described
herein.
[0033] A single-dose injection pen, or drug delivery device is
typically a disposable device which uses a sealed container which
comprises a single dose of an effective amount of a PTHrP in the
compositions described herein. A multi-dose injection pen or drug
delivery device typically contains more than one dose of an
effective amount of a PTHrP thereof in the compositions described
herein. The multi-dose pen can typically be adjusted to administer
the desired volume of the storage stable compositions described
herein. In certain embodiment the multi-dose injection pen prevents
the ingress of microbial contaminants from entering the container
or cartridge which can occur through multiple uses of one
needle.
[0034] Injection pens, as used herein, can also comprise two
containers one of which contains a PTHrP, as described herein, in a
lyophilized powder, as described below, and the second container
contains a liquid for reconstitution of the lyophilized powder. The
contents of the two containers can be mixed prior to
administration.
[0035] As discussed above the compositions of the present invention
can be administered by injection. Suitable volumes of the
compositions of the present invention for injection include about
0.5 to about 1 ml, about 0.1 to about 1 ml, about 0.02- to about
0.04 ml, about 0.1- to about 5.0 .mu.l, or about 0.1- to about 1.0
.mu.l.
[0036] In the compositions of the present invention the
concentration of the peptides is from about 20 ug/ml to about
20,000 ug/ml, from about 100 ug/ml to about 10,000 ug/ml, from
about 300 ug/ml to about 3000 ug/ml, from about 500 ug/ml to about
2000 ug/ml and about 2 mg/ml.
[0037] The compositions of the present invention can also be
lyophilized using lyophilization techniques known in the art and
stored as a powder which can be reconstituted prior to
administration. The term "lyophilization" as used herein is a
freeze drying or dehydration technique which involves removing a
solvent, preferably a water miscible solvent, more preferably water
from a composition or the present invention, typically by
sublimation under high vacuum when the composition is in a frozen
state. Typically, lyophilization is carried out in lyophilization
equipment (a lyophilizer), which comprises a drying chamber with
variable temperature controls, a condenser to collect water, and a
vacuum system to reduce the pressure in the drying chamber.
[0038] The terms "lyophilized composition", as used herein mean the
solid residue or powder which is produced or which remains after
the lyophilization procedure as defined above. The lyophilized
composition of the present invention typically further comprise a
pharmaceutically acceptable excipient. The term "pharmaceutically
acceptable excipient" as used herein refers to a substance which is
added to a solution prior to lyophilization to enhance
characteristics such as the color, texture, strength, and volume of
the lyophilized cake. Pharmaceutically acceptable excipients may
be, for example, buffers and pH adjusters, crystalline bulking
excipients, stabilizers, and tonicity raising agents.
[0039] In certain preferred embodiments the pharmaceutically
acceptable excipient is a crystalline bulking excipient. The terms
"crystalline bulking excipient" or "crystalline bulking agent" as
used herein means an excipient which provides bulk and structure to
the lyophilization cake. These crystalline bulking agents are inert
and do not react with the peptide. In addition, the crystalline
bulking agents are capable of crystallizing under lyophilization
conditions.
[0040] Examples of suitable crystalline bulking agents include
hydrophilic excipients, such as, water soluble polymers; sugars,
such as mannitol, sorbitol, xylitol, glucitol, ducitol, inositiol,
arabinitol, arabitol, galactitol, iditol, allitol, maltitol,
fructose, sorbose, glucose, xylose, trehalose, allose, dextrose,
altrose, lactose, glucose, fructose, gulose, idose, galactose,
talose, ribose, arabinose, xylose, lyxose, sucrose, maltose,
lactose, lactulose, fucose, rhamnose, melezitose, maltotriose,
raffinose, altritol, their optically active forms (D- or L-forms)
as well as the corresponding racemates; inorganic salts, both
mineral and mineral organic, such as, calcium salts, such as the
lactate, gluconate, glycerylphosphate, citrate, phosphate monobasic
and dibasic, succinate, sulfate and tartrate, as well as the same
salts of aluminum and magnesium; carbohydrates, such as, the
conventional mono- and di-saccharides as well as the corresponding
polyhydric alcohols; proteins, such as, albumin; amino acids, such
as glycine; emulsifiable fats and polyvinylpyrrolidone. Preferred
crystalline bulking agents are selected from the group consisting
of glycine, mannitol, dextran, dextrose, lactose, sucrose,
polyvinylpyrrolidone, trehalose, glucose and combinations thereof.
Particularly useful bulking agents include dextran.
[0041] As used herein a stabilizer is a composition which maintains
the chemical, biological or hormonal stability of the peptide.
Examples of stabilizing agent include polyols which includes a
saccharide, preferably a monosaccharide or disaccharide, e.g.,
glucose, trehalose, raffinose, or sucrose; a sugar alcohol such as,
for example, mannitol, sorbitol or inositol, a polyhydric alcohol
such as glycerine or propylene glycol or mixtures thereof and
albumin.
[0042] The compositions described herein can be used to stimulate
bone growth in a subject. Thus they are useful in the treatment of
diseases or disorders associated with deficiency in bone growth
such as osteoporosis and bone fractures. In one embodiment, the
present invention is a method of treating osteoporosis in a subject
comprising administering to the subject an effective amount of
composition described herein.
[0043] As used herein, "treating" can include both prophylactic,
and therapeutic treatment. For example, therapeutic treatment can
include delaying inhibiting or preventing the progression of
osteoporosis, the reduction or elimination of symptoms associated
with osteoporosis. Prophylactic treatment can include preventing,
inhibiting or delaying the onset of osteoporosis.
[0044] As used herein, an effective amount refers to an amount
sufficient to elicit the desired response. In the present
invention, the desired biological response is a decrease in the
rate of bone loss and/or an increase in the bone mass or bone
quality of a subject.
[0045] Suitable dosage for use in the compositions and methods of
the present invention include from about 40 to about 160 .mu.g,
about 80 to about 120 .mu.g about 80 to about 100 .mu.g; or from
about 40 to about 50 .mu.g, about 50 to about 60 .mu.g, about 60 to
about 70 .mu.g, about 70 to about 80 .mu.g, about 80 to about 90
.mu.g, about 90 to about 100 .mu.g, about 100 to about 110 .mu.g,
about 110 to about 120 .mu.g, about 120 to about 130 .mu.g, about
130 to about 140 .mu.g, about 140 to about 150 .mu.g, about 150 to
about 160 .mu.g; or from 40 to about 45 .mu.g, about 45 to about 50
.mu.g, about 50 to about 55 .mu.g, about 55 to about 60 .mu.g,
about 60 to about 65 .mu.g, about 65 to about 70 .mu.g, about 70 to
about 75 .mu.g, about 75 to about 80 .mu.g, about 80 to about 85
.mu.g, about 85 to about 90 .mu.g, about 90 to about 95 .mu.g,
about 95 to about 100 .mu.g, about 100 to about 105 .mu.g, about
105 to about 110 .mu.g, about 110 to about 115 .mu.g, about 115 to
about 120 .mu.g, about 120 to about 125 .mu.g, about 125 to about
130 .mu.g, about 130 to about 135 .mu.g, about 135 to about 140
.mu.g, about 140 to about 145 .mu.g, about 145 to about 150 .mu.g,
about 150 to about 155 .mu.g, about 155 to about 160 .mu.g
administered once per day, once every other day, twice per week
once per week, once every two weeks, once per month. The doses can
be a pulsatile injection, for example, once per month which causes
pulsatile release of singles doses of the composition described
herein.
[0046] When the dosages described above are administered once per
day, once per week etc., typically the dosages are of equal
amounts.
[0047] The subject as used herein can be an animal, for example, a
mammal, such as a human.
[0048] In certain embodiments of this invention, compositions
comprising dosage forms containing 20 .mu.g, 40 .mu.g, or 80 .mu.g
of [Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2(SEQ ID NO.: 2) are
described.
[0049] In certain embodiments of this invention, a method of
treatment of osteoporosis is described wherein doses of 20 .mu.g,
40 .mu.g, or 80 .mu.g of [Glu.sup.22,25, Leu.sup.23,28,31,
Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) are
administered by daily subcutaneous injection to a subject in need
thereof.
[0050] In some embodiments, the subject in need thereof has
osteoporosis.
[0051] In some embodiments, the subject in need thereof has
osteopenia.
[0052] In certain embodiments, the subject in need thereof is a
post-menopausal woman.
[0053] In some embodiments, the subject in need thereof has
glucocorticoid induced osteoporosis.
[0054] In certain embodiments, the subject in need thereof has
glucocorticoid induced osteopenia.
[0055] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in plasma Cmax levels of SEQ ID NO. 2 between 32.4 pg/mL and 53.8
pg/mL.
[0056] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in plasma Cmax levels of SEQ ID NO. 2 between 32.4 pg/mL
and 53.8 pg/mL.
[0057] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in plasma Cmax levels of SEQ ID NO. 2 between 61.1 pg/mL and 168.9
pg/mL.
[0058] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in plasma Cmax levels of SEQ ID NO. 2 between 61.1 pg/mL
and 168.9 pg/mL.
[0059] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in plasma Cmax levels of SEQ ID NO. 2 between 124 pg/mL and 322
pg/mL.
[0060] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 uG SEQ ID NO. 2
resulting in plasma Cmax levels of SEQ ID NO. 2 between 124 pg/mL
and 322 pg/mL.
[0061] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in Cmax plasma levels of SEQ ID NO. 2 between 255.57 pg/mL and
364.3 pg/mL.
[0062] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in Cmax plasma levels of SEQ ID NO. 2 between 255.57
pg/mL and 364.3 pg/mL.
[0063] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.531 hours and 1.00
hours.
[0064] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.531
hours and 1.00 hours.
[0065] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.250 hours and
0.624 hours.
[0066] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.250
hours and 0.624 hours.
[0067] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.262 hours and
0.579 hours.
[0068] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.262
hours and 0.579 hours.
[0069] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.251 hours and 1.01
hours.
[0070] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.251
hours and 1.01 hours.
[0071] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.90 hours and 3.28
hours.
[0072] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.90 hours
and 3.28 hours.
[0073] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 0.736 hours and 1.364
hours.
[0074] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 0.736 hours
and 1.364 hours.
[0075] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.396 hours and 1.904
hours.
[0076] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.396 hours
and 1.904 hours.
[0077] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.585 hours and 3.015
hours.
[0078] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.585 hours
and 3.015 hours.
[0079] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between 132.82 pg
h/mL and 241.90 pg h/mL.
[0080] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between
132.82 pg h/mL and 241.90 pg h/mL.
[0081] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between 138.12 pg
h/mL and 376.22 pg h/mL.
[0082] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between
138.12 pg h/mL and 376.22 pg h/mL.
[0083] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between 311.54 pg
h/mL and 874.34 pg h/mL.
[0084] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between
311.54 pg h/mL and 874.34 pg h/mL.
[0085] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between 541.99 pg
h/mL and 1569.21 pg h/mL.
[0086] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-inf) of SEQ ID NO. 2 between
541.99 pg h/mL and 1569.21 pg h/mL.
[0087] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 33.2 pg/mL and 48.4
pg/mL.
[0088] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 33.2 pg/mL
and 48.4 pg/mL.
[0089] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 89.8 pg/mL and 128.2
pg/mL.
[0090] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 89.8 pg/mL
and 128.2 pg/mL.
[0091] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 129.3 pg/mL and
284.4 pg/mL.
[0092] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 uG SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 129.3
pg/mL and 284.4 pg/mL.
[0093] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 367.2 pg/mL and
504.8 pg/mL.
[0094] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 367.2
pg/mL and 504.8 pg/mL.
[0095] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.514 hours and 1.53
hours.
[0096] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.514
hours and 1.53 hours.
[0097] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.250 hours and 3.05
hours.
[0098] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.250
hours and 3.05 hours.
[0099] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.349 hours and 1.00
hours.
[0100] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.349
hours and 1.00 hours.
[0101] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma T.sub.max for SEQ ID NO. 2 between 0.500 hours and 1.00
hours.
[0102] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a plasma T.sub.max for SEQ ID NO. 2 between 0.500
hours and 1.00 hours.
[0103] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 0.806 hours and 1.294
hours.
[0104] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 0.806 hours
and 1.294 hours.
[0105] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.033 hours and 1.827
hours.
[0106] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.033 hours
and 1.827 hours.
[0107] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.265 hours and 2.115
hours.
[0108] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a plasma t.sub.1/2 of SEQ ID NO. 2 between 1.265 hours
and 2.115 hours.
[0109] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-2.5h) of SEQ ID NO. 2 between 50.263 pg
h/mL and 111.14 pg h/mL.
[0110] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-2.5h) of SEQ ID NO. 2 between
50.263 pg h/mL and 111.14 pg h/mL.
[0111] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-3.0h) of SEQ ID NO. 2 between 89.549 pg
h/mL and 253.611 pg h/mL.
[0112] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-3.0h) of SEQ ID NO. 2 between
89.549 pg h/mL and 253.611 pg h/mL.
[0113] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-3.49h) of SEQ ID NO. 2 between 188.28 pg
h/mL and 627.68 pg h/mL.
[0114] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-3.49h) of SEQ ID NO. 2 between
188.28 pg h/mL and 627.68 pg h/mL.
[0115] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a net plasma AUC.sub.(0-6.00h) of SEQ ID NO. 2 between 619.55 pg
h/mL and 1386.45 pg h/mL.
[0116] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g of SEQ ID NO. 2
resulting in a net plasma AUC.sub.(0-6.00h) of SEQ ID NO. 2 between
619.55 pg h/mL and 1386.45 pg h/mL.
[0117] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 32.4 pg/mL and 53.8 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.531 hours and 1.00 hours.
[0118] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level between 61.1 pg/mL and 168.9 pg/mL of SEQ ID
NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between 0.25 hours
and 0.624 hours.
[0119] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 61.1 pg/mL and 168.9 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.25 hours and 0.624 hours.
[0120] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 124 pg/mL and 322
pg/mL and a plasma T.sub.max for SEQ ID NO. 2 for SEQ ID NO. 2
between 0.262 hours and 0.579 hours.
[0121] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 124 pg/mL
and 322 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 between 0.262
hours and 0.579 hours.
[0122] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a Cmax plasma level of SEQ ID NO. 2 between 255.57 pg/mL and
364.3 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 of between
0.251 hours and 1.01 hours.
[0123] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a Cmax plasma level between 255.57 pg/mL and 364.3
pg/mL and a plasma T.sub.max for SEQ ID NO. 2 between 0.251 hours
and 1.01 hours.
[0124] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 33.2 pg/mL and 48.4 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.514 hours and 1.53 hours.
[0125] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level between 89.8 pg/mL and 128.2 pg/mL of SEQ ID
NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between 0.250 hours
and 3.05 hours.
[0126] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 89.8 pg/mL and 128.2 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.250 hours and 3.05 hours.
[0127] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 129.3 pg/mL and
284.4 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 for SEQ ID NO.
2 between 0.349 hours and 1.00 hours.
[0128] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 129.3
pg/mL and 284.4 pg/mL and a plasma T.sub.max for SEQ ID NO. 2
between 0.349 hours and 1.00 hours.
[0129] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a Cmax plasma level of SEQ ID NO. 2 between 367.2 pg/mL and
504.8 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 of between
0.500 hours and 1.00 hours.
[0130] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a Cmax plasma level between 367.2 pg/mL and 504.8
pg/mL and a plasma T.sub.max for SEQ ID NO. 2 between 0.500 hours
and 1.00 hours.
[0131] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 5 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 32.4 pg/mL and 53.8 pg/mL
of SEQ ID NO. 2, a plasma T.sub.max for SEQ ID NO. 2 between 0.531
hours and 1.00 hours and a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 1.90 and 3.28 hours.
[0132] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level between 61.1 pg/mL and 168.9 pg/mL of SEQ ID
NO. 2, a plasma T.sub.max for SEQ ID NO. 2 between 0.25 hours and
0.624 hours and a plasma t.sub.1/2 for SEQ ID NO. 2 of between
0.736 hours and 1.364 hours.
[0133] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 61.1 pg/mL and 168.9 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.25 hours and 0.624 hours and a plasma t.sub.1/2 for SEQ ID NO. 2
of between 0.736 hours and 1.364 hours.
[0134] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 124 pg/mL and 322
pg/mL and a plasma T.sub.max for SEQ ID NO. 2 for SEQ ID NO. 2
between 0.262 hours and 0.579 hours and a plasma t.sub.1/2 for SEQ
ID NO. 2 of between 1.396 hours and 1.904 hours.
[0135] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 124 pg/mL
and 322 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 between 0.262
hours and 0.579 hours and a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 0.736 hours and 1.364 hours.
[0136] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a Cmax plasma level of SEQ ID NO. 2 between 255.57 pg/mL and
364.3 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 of between
0.251 hours and 1.01 hours and a plasma t.sub.1/2 for SEQ ID NO. 2
of between 1.585 hours and 3.015 hours.
[0137] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a Cmax plasma level between 255.57 pg/mL and 364.3
pg/mL and a plasma T.sub.max for SEQ ID NO. 2 between 0.251 hours
and 1.01 hours and a plasma t.sub.1/2 for SEQ ID NO. 2 of between
1.585 hours and 3.015 hours.
[0138] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level between 89.8 pg/mL and 128.2 pg/mL of SEQ ID
NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between 0.250 hours
and 3.05 hours and a plasma t.sub.1/2 for SEQ ID NO. 2 of between
0.806 hours and 1.294 hours.
[0139] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 89.8 pg/mL and 128.2 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.250 hours and 3.05 hours and a plasma t.sub.1/2 for SEQ ID NO. 2
of between 0.806 hours and 1.294 hours.
[0140] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 129.3 pg/mL and
284.4 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 for SEQ ID NO.
2 between 0.349 hours and 1.00 hours and a plasma t.sub.1/2 for SEQ
ID NO. 2 of between 1.033 hours and 1.827 hours.
[0141] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 129.3
pg/mL and 284.4 pg/mL and a plasma T.sub.max for SEQ ID NO. 2
between 0.349 hours and 1.00 hours and a plasma t.sub.1/2 for SEQ
ID NO. 2 of between 1.033 hours and 1.827 hours.
[0142] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a Cmax plasma level of SEQ ID NO. 2 between 367.2 pg/mL and
504.8 pg/mL and a plasma T.sub.max for SEQ ID NO. 2 of between
0.500 hours and 1.00 hours and a plasma t.sub.1/2 for SEQ ID NO. 2
of between 1.265 hours and 2.115 hours.
[0143] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a Cmax plasma level between 367.2 pg/mL and 504.8
pg/mL and a plasma T.sub.max for SEQ ID NO. 2 between 0.500 hours
and 1.00 hours and a plasma t.sub.1/2 for SEQ ID NO. 2 of between
1.265 hours and 2.115 hours.
[0144] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level between 61.1 pg/mL and 168.9 pg/mL of SEQ ID
NO. 2, a plasma T.sub.max for SEQ ID NO. 2 between 0.25 hours and
0.624 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of between 0.736
hours and 1.364 hours and a net plasma AUC.sub.(0-inf) of SEQ ID
NO. 2 between 138.12 pg h/mL and 376.22 pg h/mL.
[0145] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 61.1 pg/mL and 168.9 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.25 hours and 0.624 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 0.736 hours and 1.364 hours and a net plasma
AUC.sub.(0-inf) of SEQ ID NO. 2 between 138.12 pg h/mL and 376.22
pg h/mL.
[0146] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 124 pg/mL and 322
pg/mL, a plasma T.sub.max for SEQ ID NO. 2 for SEQ ID NO. 2 between
0.262 hours and 0.579 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 1.396 hours and 1.904 hours and a net plasma
AUC.sub.(0-inf) of SEQ ID NO. 2 between 311.54 pg h/mL and 874.34
pg h/mL.
[0147] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 124 pg/mL
and 322 pg/mL, a plasma T.sub.max for SEQ ID NO. 2 between 0.262
hours and 0.579 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 0.736 hours and 1.364 hours and a net plasma
AUC.sub.(0-inf) of SEQ ID NO. 2 between 311.54 pg h/mL and 874.34
pg h/mL.
[0148] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a Cmax plasma level of SEQ ID NO. 2 between 255.57 pg/mL and
364.3 pg/mL, a plasma T.sub.max for SEQ ID NO. 2 of between 0.251
hours and 1.01 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 1.585 hours and 3.015 hours and a net plasma
AUC.sub.(0-inf) of SEQ ID NO. 2 between 311.54 pg h/mL and 874.34
pg h/mL.
[0149] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a Cmax plasma level between 255.57 pg/mL and 364.3
pg/mL, a plasma T.sub.max for SEQ ID NO. 2 between 0.251 hours and
1.01 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of between 1.585
hours and 3.015 hours and a net plasma AUC.sub.(0-inf) of SEQ ID
NO. 2 between 541.99 pg h/mL and 1569.21 pg h/mL.
[0150] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level between 89.8 pg/mL and 128.2 pg/mL of SEQ ID
NO. 2, a plasma T.sub.max for SEQ ID NO. 2 between 0.250 hours and
3.05 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of between 0.806
hours and 1.294 hours and a net plasma AUC.sub.(0-3.00h) of SEQ ID
NO. 2 between 89.549 pg h/mL and 253.611 pg h/mL.
[0151] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 20 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level between 89.8 pg/mL and 128.2 pg/mL
of SEQ ID NO. 2 and a plasma T.sub.max for SEQ ID NO. 2 between
0.250 hours and 3.05 hours and a plasma t.sub.1/2 for SEQ ID NO. 2
of between 0.806 hours and 1.294 hours and a net plasma
AUC.sub.(0-3.00h) of SEQ ID NO. 2 between 89.549 pg h/mL and
253.611 pg h/mL.
[0152] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a plasma Cmax level of SEQ ID NO. 2 between 129.3 pg/mL and
284.4 pg/mL, a plasma T.sub.max for SEQ ID NO. 2 for SEQ ID NO. 2
between 0.349 hours and 1.00 hours, a plasma t.sub.1/2 for SEQ ID
NO. 2 of between 1.033 hours and 1.827 hours and a net plasma
AUC.sub.(0-3.49h) of SEQ ID NO. 2 between 188.28 pg h/mL and 627.68
pg h/mL.
[0153] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 40 .mu.g SEQ ID NO. 2
resulting in a plasma Cmax level of SEQ ID NO. 2 between 129.3
pg/mL and 284.4 pg/mL, a plasma T.sub.max for SEQ ID NO. 2 between
0.349 hours and 1.00 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 1.033 hours and 1.827 hours and a net plasma
AUC.sub.(0-3.49h) of SEQ ID NO. 2 between 188.28 pg h/mL and
627.681 pg h/mL.
[0154] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of SEQ ID NO. 2 resulting
in a Cmax plasma level of SEQ ID NO. 2 between 367.2 pg/mL and
504.8 pg/mL, a plasma T.sub.max for SEQ ID NO. 2 of between 0.500
hours and 1.00 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of
between 1.265 hours and 2.115 hours and a net plasma
AUC.sub.(0-6.00h) of SEQ ID NO. 2 between 619.55 pg h/mL and
1386.45 pg h/mL.
[0155] In some embodiments this invention provides a method of
treatment of osteoporosis comprising treating a subject in need
thereof by daily subcutaneous injection of 80 .mu.g SEQ ID NO. 2
resulting in a Cmax plasma level between 367.2 pg/mL and 504.8
pg/mL, a plasma T.sub.max for SEQ ID NO. 2 between 0.500 hours and
1.00 hours, a plasma t.sub.1/2 for SEQ ID NO. 2 of between 1.265
hours and 2.115 hours and a net plasma AUC.sub.(0-6.00h) of SEQ ID
NO. 2 between 619.55 pg h/mL and 1386.45 pg h/mL.
[0156] For purposes of this invention, Cmax refers to the maximum
concentration that is measured in the plasma. Tmax is the time at
which the maximum concentration occurs. AUC refers to the integral
area under the curve for a given time interval. AUC.sub.(0-inf)
refers to the area under the curve as extrapolated out to infinity
or (maximum area under the curve). AUC.sub.(0-t) refers to the area
under the curve at the time in hours listed. For example,
AUC.sub.(0-7h) indicates the area under the curve after 7 hours.
t.sub.1/2 refers to the 1/2 life of the drug.
[0157] For the purposes of this disclosure, the therapeutic utility
of these compounds includes "treating" a human and methods of
treatment or treating a subject, human or patient, where treating
is understood to include treating, preventing, or ameliorating the
symptoms associated with, or reducing the incidence of, reducing
the pathogenesis of, facilitating the recovery from or delaying the
onset of the syndrome, illness, malady or condition being
considered. As it pertains to osteoporosis and the methods of this
invention, a method of treatment should also be understood to
include a method of preventing osteoporosis. Increasing bone
mineral density in a population with osteoporosis can accordingly
be deemed a treatment for osteoporosis in that patient population.
Likewise, preventing osteoporosis can be accomplished by
administering the compositions and compounds of this invention to a
patient population that does not yet have osteoporosis. In some
embodiments of this invention, the patient population being
administered the compositions and/or according to the methods of
this invention are at increased risk for osteoporosis or who
already have osteoporosis. It should also be appreciated that
osteopenia is included with osteoporosis for purposes of this
invention.
[0158] One of skill in the art appreciates that the several
disorders are associated with osteoporosis and so it should be
appreciated that the methods and compositions of this invention are
useful for treating osteoporosis from the many origins and risk
factors from which osteoporosis and osteopenia arise including but
not limited to osteogenesis imperfecta, Mafan syndrome,
hemochromatosis, hypophosphatasia, glycogen storage diseases,
homocysinuria, Ehlers-Danlos syndrome, porhyria, Menke's syndrome,
epidermolysis bullosa and Gaucher's disease.
[0159] A pharmaceutically acceptable salt is a salt which is
suitable for administration to a subject, such as, a human. The
peptides of the present invention can have one or more sufficiently
acidic proton that can react with a suitable organic or inorganic
base to form a base addition salt. Base addition salts include
those derived from inorganic bases, such as ammonium or alkali or
alkaline earth metal hydroxides, carbonates, bicarbonates, and the
like, and organic bases such as alkoxides, alkyl amides, alkyl and
aryl amines, and the like. Such bases useful in preparing the salts
of this invention thus include sodium hydroxide, potassium
hydroxide, ammonium hydroxide, potassium carbonate, and the like.
The peptides of the present invention having a sufficiently basic
group, such as an amine can react with an organic or inorganic acid
to form an acid addition salt. Acids commonly employed to form acid
addition salts from compounds with basic groups are inorganic acids
such as hydrochloric acid, hydrobromic acid, hydroiodic acid,
sulfuric acid, phosphoric acid, and the like, and organic acids
such as p-toluenesulfonic acid, methanesulfonic acid, oxalic acid,
p-bromophenyl-sulfonic acid, carbonic acid, succinic acid, citric
acid, benzoic acid, acetic acid, and the like. Examples of such
salts include the sulfate, pyrosulfate, bisulfate, sulfite,
bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate,
metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate,
propionate, decanoate, caprylate, acrylate, formate, isobutyrate,
caproate, heptanoate, propiolate, oxalate, malonate, succinate,
suberate, sebacate, fumarate, maleate, butyne-1,4-dioate,
hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate,
dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate,
sulfonate, xylenesulfonate, phenylacetate, phenylpropionate,
phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate,
tartrate, methanesulfonate, propanesulfonate,
naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and
the like.
[0160] The compositions of the present invention typically do not
show any or show reduced side-effects such as hypercalcemia and
typically do not increase the stimulation of bone resorption at the
dosage listed above. This reduction in side effects allows for
administration of higher doses than commercially available
osteoporosis drugs.
[0161] The compositions of the present invention can be
administered by injection as described herein or by pulmonary or
transdermal delivery.
[0162] The compositions of the present invention may be
administered alone or in combination with an additional therapeutic
agent, such as an antiresorptive therapy, for example,
bisphonsphonates and calcitonin.
EXEMPLIFICATION
EXAMPLE 1
Demonstrates Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,38]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) Stability at Low
Acetate Concentration (1 mM), without Stabilizer
TABLE-US-00002 [0163] TABLE 1 Unitary Formula Material Supplier
(per cartridge) (SEQ ID NO.: 2) Ipsen Ireland 0.140 mg (free base)
Tri-hydrate sodium acetate 0.1N Prolabo 14.6 mg Acetic acid 0.1N
Prolabo 1.9 mg qs pH 5.1 Water for Injection Meram qs 1.4 g Type I
clear glass Cartridge 1.5 ml, Bunderglass 1 washed, siliconised and
sterilised via Vetter Grey PTFE bromobutyl Daikyo 1 cartridge
rubber stopper Chlorobutyl rubber-metal West 1 cartridge crimp
Pharmaceutical qs = quantity sufficient to achieve
[0164] The formulation delivered 100 mcg of (SEQ ID NO.: 2) per 0.1
ml. (SEQ ID NO.: 2) was dissolved in Water for Injection containing
dilute acetate buffer to give pH 5.1 was used.
[0165] Results confirm excellent chemical stability over 24 months,
at 5.degree. C. as shown in FIG. 1. This solution contains no
stabilizer or preservative and only 6 mM acetate buffer.
[0166] In summation for (SEQ ID NO.: 2), stabilizer is not needed
to give good stability in solution.
EXAMPLE 2
Use of Citric Acid Buffer in Lyophilised Form of (SEQ ID NO.:
2)
TABLE-US-00003 [0167] TABLE 2 Unitary Material Supplier Formula
(per vial) (SEQ ID NO.: 2) Ipsen Ireland 0.1 mg (free base) Dextran
70 Interchemical 50 mg Citric acid 0.25% (w/v) Prolabo qs pH 4.5*
Water for injections** Meram qs 1 g Type I clear glass vial, 11-13
ml Verretubex 1 Grey chlorobutyl PTFE stopper, Daikyo 1 20 mm West
1 Flip-off metal crimp Pharma **to get pH 5-5.5 after
lyophilisation removed after freeze-drying step.
[0168] The solutions in Table 2 were reconstituted with NaCl 0.9%,
to give:
[0169] ONE vial of 2 ml (=50 .mu.g/ml) providing 10 to 80 .mu.g/d
doses (with injections of 200 .mu.l to 1.6 ml), or
[0170] ONE vial of 5 ml (=20 .mu.g/ml solution) providing 5 to 40
.mu.g/d doses (with injections of 250 .mu.l-2 ml).
[0171] Citric acid was used to adjust pH and Dextran was used to
provide a bulking agent to aid cake formation during
lyophilization.
[0172] The solutions described were lyophilized in glass vials, and
stored at various temperatures for up to 24 months. The content of
Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2(SEQ ID NO.: 2), purity and
physical tests were conducted on samples removed from storage at
different times. Results are presented in FIG. 2, for peptide
concentration, as percent remaining The data in FIG. 2 shows
excellent stability over 24 months at 2-8.degree. C.
EXAMPLE 3
Screening of Formulations for Glu.sup.22,25, Leu.sup.23,28,31,
Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2(SEQ ID NO.: 2) to
Compare Different Preservatives
[0173] TABLE 3 below shows Methyparaben and Benzyl Alcohol are not
suitable preservatives for use with Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2), as precipitation and/or inactivity in preservative
activity was seen.
TABLE-US-00004 TABLE 3 Example 3a Example 3b Example 3c Example 3d
Example 3e Methylparaben 1.5 mg/mL 1.35 mg/mL -- -- --
Propylparaben -- 0.15 mg/mL -- -- -- Phenol -- -- 5 mg/mL -- --
Chlorocresol -- -- -- 3 mg/mL -- Benzyl alcohol -- -- -- -- 10
mg/mL Preservative Failed Pass Pass Pass Pass effectiveness test
Observationo or Precipitation -- -- -- -- Issues observed
Preservative Not Tested Pass Pass Pass Fail effectiveness as test
after storage precipitated 4.5 months at 5.degree. C. initially
[0174] Solutions were prepared containing Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2) 2 mg/ml, acetate buffer 6 mM and water for
injection, with various different preservatives added at
concentrations recommended for effective antimicrobial activity.
Solutions were prepared at room temperature, by dissolution of the
various ingredients in water for injection, with stirring over
<30 minutes to ensure complete dissolution, Solutions were
filtered thro.mu.gh 0.2 micron filter and filled into glass vials,
to which a rubber stopper was applied and crimped in place to
ensure complete closure.
[0175] The solution with methylparaben was unacceptable due to
precipitation and inactivity immediately after manufacture of the
solution. The solutions were then stored for up to 3 months at
25.degree. C., and up to 4.5 months at 5.degree. C. and the
preservative effectiveness test repeated. as described in Example
5.
EXAMPLE 4
Evaluation of Anti-Microbial Preservative Effectiveness of Various
Concentrations of Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) Compositions
(Stability Study)
TABLE-US-00005 [0176] TABLE 4 P87228 P87229 P87230 P87231 (SEQ ID 2
mg/mL 2 mg/mL 2 mg/mL 2 mg/mL NO.: 2) Anti-microbial Phenol
Chlorocresol Chlorocresol Benzyl alcohol 5 mg/mL 3 mg/mL 2 mg/mL 10
mg/mL Acetate buffer pH 5.1 pH 5.1 pH 5.1 pH 5.1
[0177] The solutions were tested according to European
Pharmacopoeia, Chapter 5.1.3 "Efficacite de la conservation
anti-microbienne" (Anti-microbial effectiveness test) to prove the
effectiveness of the preservative.
TABLE-US-00006 TABLE 5 Preservative effectiveness test after
manufacturing Nb of cfu present/preparation Initial (plate-count
method) organism P87228 P87229 P87230 P87231 Organisms:
concentration Test Phenol Chlorocresol Chlorocresol Benzyl alc.
Bacteria in cfu/mL interval 5 mg/ml 3 mg/ml 2 mg/ml 10 mg/ml
Staphylococcus 3.8 .times. 10.sup.5 T 0 3.4 .times. 10.sup.5 <5
<5 4.7 .times. 10.sup.5 aureus T + 6 hrs <5 <5 <5 6.8
.times. 10.sup.2 T + 24 hrs <5 <5 <5 <5 T + 28 5 (*)
<5 <5 <5 days Pseudomonas 1.3 .times. 10.sup.6 T 0 5 <5
<5 1.5 .times. 10.sup.2 aeruginosa T + 6 hrs <5 <5 <5
<5 T + 24 hrs <5 <5 <5 <5 T + 28 <5 <5 <5
<5 days E. coli 6.7 .times. 10.sup.5 T 0 7.2 .times. 10.sup.3
<5 <5 1.1 .times. 10.sup.5 T + 6 hrs <5 <5 <5 <5
T + 24 hrs <5 <5 <5 <5 T + 28 <5 <5 <5 <5
days Nb of cfu present/preparation Initial (plate-count method)
Organism: organism P87228 P87229 P87230 P87231 Yeast and
concentration Test Phenol chlorocresol chlorocresol Benzyl alc.
mold in cfu/mL interval 5 mg/ml 3 mg/ml 2 mg/ml 10 mg/ml
Aspergillus 3.4 .times. 10.sup.5 T 0 4.0 .times. 10.sup.5 <5
<5 4.1 .times. 10.sup.5 niger T + 7 <5 <5 <5 <5 days
T + 28 <5 <5 <5 <5 days Candida 3.9 .times. 10.sup.5 T
0 4.4 .times. 10.sup.5 <5 <5 3.8 .times. 10.sup.5 albicans T
+ 7 <5 <5 <5 5 days T + 28 <5 <5 <5 <5 days
Results: Conform -- Conform Conform Conform Conform Preservative
effectiveness test results after 3 months storage at 25.degree. C.
Nb of cfu present/preparation Initial (plate-count method) organism
Test P87228 P87229 P87230 P87231 Organisms: concentration interval
Phenol chlorocresol chlorocresol Benzyl alc. Bacteria in cfu/mL
(days) 5 mg/ml 3 mg/ml 2 mg/ml 10 mg/ml Staphylococcus 2.7 .times.
10.sup.5 0 hr 1.9 .times. 10.sup.5 <5 <5 3.8 .times. 10.sup.5
aureus (P87228, 6 hr 30 <5 <5 5.9 .times. 10.sup.3 P87229,
P87231) 24 hr <5 <5 <5 <5 5.2 .times. 10.sup.5 28 day
<5 <5 <5 <5 (P87230) Pseudomonas 9.9 .times. 10.sup.5 0
hr <5 <5 <5 <5 aeruginosa (P87228, 6 hr <5 <5
<5 <5 P87229, P87231) 24 hr <5 <5 <5 <5 8.5
.times. 10.sup.5 28 day <5 <5 <5 <5 (P87230) E. coli
6.8 .times. 10.sup.5 0 hr 1.7 .times. 10.sup.5 <5 <5 8.0
.times. 10.sup.4 (P87228, 6 hr <5 <5 <5 5 P87229, P87231)
24 hr <5 <5 <5 <5 9.5 .times. 10.sup.5 28 day <5
<5 <5 <5 (P87230) Nb of cfu present/preparation Initial
(plate-count method) Organism: organism P87228 P87229 P87230 P87231
Yeast and concentration Test Phenol Chlorocresol Chlorocresol
Benzyl alc. mold in cfu/mL interval 5 mg/ml 3 mg/ml 2 mg/ml 10
mg/ml Aspergillus 3.3 .times. 10.sup.5 0 hr 3.8 .times. 10.sup.5 55
70 4.1 .times. 10.sup.5 niger (P87228, 7 day <5 <5 <5
<5 P87229, P87231) 28 day <5 <5 <5 <5 4.1 .times.
10.sup.5 (P87230) Candida 2.7 .times. 10.sup.5 0 hr 4.0 .times.
10.sup.5 <5 <5 3.8 .times. 10.sup.5 albicans (P87228, 7 day
<5 <5 <5 <5 P87229, P87231) 28 day <5 <5 <5
<5 3.7 .times. 10.sup.5 (P87230) Results: Conform -- Conform
Conform Conform Not Conform Preservative effectiveness test results
after 4.5 months storage at 5.degree. C. Nb of cfu
present/preparation Initial (plate-count method) organism Test
P87228 P87229 P87230 P87231 Organisms: concentration interval
Phenol chlorocresol chlorocresol Benzyl alc. Bacteria in cfu/mL
(days) 5 mg/ml 3 mg/ml 2 mg/ml 10 mg/ml Staphylococcus 5.4 .times.
10.sup.5 0 hr 4.1 .times. 10.sup.5 <5 <5 5.1 .times. 10.sup.5
aureus 6 hr <5 <5 <5 7.2 .times. 10.sup.3 24 hr <5
<5 <5 <5 28 day <5 <5 <5 <5 Pseudomonas 9.7
.times. 10.sup.5 0 hr <5 <5 <5 <5 aeruginosa 6 hr <5
<5 <5 <5 24 hr <5 <5 <5 <5 28 day <5 <5
<5 <5 E. coli 6.1 .times. 10.sup.5 0 hr 7.0 .times. 10.sup.4
5 5 4.2 .times. 10.sup.4 6 hr <5 <5 <5 <5 24 hr <5
<5 <5 <5 28 day <5 <5 <5 <5 Nb of cfu
present/preparation Initial (plate-count method) Organism: organism
P87228 P87229 P87230 P87231 Yeast and concentration Test Phenol
Chlorocresol Chlorocresol Benzyl alc. mold in cfu/mL interval 5
mg/ml 3 mg/ml 2 mg/ml 10 mg/ml Aspergillus 5.3 .times. 10.sup.5 0
hr 3.7 .times. 10.sup.5 1.8 .times. 10.sup.3 7.5 .times. 10.sup.3
4.1 .times. 10.sup.5 niger 7 day <5 <5 <5 <5 28 day
<5 <5 <5 <5 Candida 4.1 .times. 10.sup.5 0 hr 4.5
.times. 10.sup.5 <5 5 4.5 .times. 10.sup.5 albicans 7 day <5
<5 <5 <5 28 day <5 <5 <5 <5 Results: Conform
-- Conform Conform Conform Not Conform (*) Bacillus Gram +,
different from St. Aureus -> result conform Nb of cfu = number
of colony forming units
[0178] TABLE 5 shows Phenol, Chlorocresol and Benzyl Alcohol all
produce compliant results immediately after manufacture for both
Bacteria and Yeasts/moulds. After 3 and 4.5 months storage, the
preservative efficacy is maintained for Phenol and Chlorocresol,
for both Bacteria and Yeasts/moulds. However, for Benzyl Alcohol,
the efficacy against Bacteria is not compliant, as the data shows
insufficient rate of kill against S Aureus (TABLE 5).
EXAMPLE 5
Chemical Stability of Different Formulations
[0179] Table 6 details the chemical stability of the formulations
described in Example 4.
TABLE-US-00007 TABLE 6 Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) stability
results Storage conditions: 25.degree. C., 60% RH (SEQ ID NO.: 2)
content in mg/mL (% initial concentration at t = 0) Batch
Composition 0 month 1 month 3 months P87228 (SEQ ID NO.: 2) (2
mg/ml)/ 1.90 1.88 1.83 Phenol (5 mg/ml) (100%) (98.9%) (96.3%)
P87229 (SEQ ID NO.: 2) (2 mg/ml)/ 1.98 1.96 1.94 Chlorocresol (3
mg/ml) (100%) (99.0%) (98.0%) P87231 (SEQ ID NO.: 2) (2 mg/ml)/
1.93 1.89 1.86 Benzyl Alcohol (100%) (97.9%) (96.4%) (10 mg/ml)
Storage conditions: 5.degree. C. (SEQ ID NO.: 2) content in mg/mL
(% initial concentration at t = 0) Batch Composition 0 month 3
month 4.5 month P87228 (SEQ ID NO.: 2) 1.90 1.91 1.89 (2
mg/ml)/Phenol (100%) (100.5%) (99.5%) (5 mg/ml) P87229 (SEQ ID NO.:
2) 1.98 1.96 1.97 (2 mg/ml)/Chlorocresol (100%) (99.0%) (99.5%) (3
mg/ml) P87231 (SEQ ID NO.: 2) 1.93 1.94 1.92 (2 mg/ml)/Benzyl
Alcohol (100%) (100.5%) (99.5%) (10 mg/ml)
[0180] As can be seen from TABLE 6 and Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2(SEQ ID NO.: 2) solution
stability is not significantly influenced by the preservative
selected. TABLE 7 details the content of each preservative for the
same formulations.
TABLE-US-00008 TABLE 7 Preservative stability results Storage
conditions: 25.degree. C., 60% RH Preservative content in mg/ml (%
initial concentration at t = 0) Batch Composition 0 month 1 month 3
month P87228 (SEQ ID NO.: 2) (2 mg/ml)/ 4.86 4.82 4.79 Phenol (5
mg/ml) (100%) (99.2%) (98.6%) P87229 (SEQ ID NO.: 2) (2 mg/ml)/
2.78 2.70 2.56 Chlorocresol (3 mg/ml) (100%) (97.1%) (92.1%) P87231
(SEQ ID NO.: 2) (2 mg/ml)/ 9.92 9.83 9.82 Benzyl Alcohol (100%)
(99.1%) (99.0%) (10 mg/ml) Storage conditions: 5.degree. C.
Preservative content in mg/mL (% initial concentration at t = 0)
Batch Composition 0 month 3 month 4.5 month P87228 (SEQ ID NO.: 2)
(2 mg/ml)/ 4.86 4.83 4.84 Phenol (5 mg/ml) (100%) (99.4%) (99.6%)
P87229 (SEQ ID NO.: 2) (2 mg/ml)/ 2.78 2.73 2.74 Chlorocresol (3
mg/ml) (100%) (98.2%) (98.6%) P87231 (SEQ ID NO.: 2) (2 mg/ml)/
9.92 9.89 9.94 Benzyl Alcohol (100%) (99.7%) (100.2%) (10
mg/ml)
[0181] As can be seen from TABLE 7 chlorocresol is the preservative
which has the lower stability, with greater loss in preservative
content under both 5 and 25.degree. C. storage.
EXAMPLE 6
Clinical Study of Subjects Treated with Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2)
[0182] A randomized, double-blind, placebo-controlled,
multiple-dose design study of Glu.sup.22,25, Leu.sup.23,28,31,
Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) was
conducted at 2 sites. A total of 39 eligible subjects were
sequentially enrolled into 1 of 4 study groups consisting of 10
subjects each, with the exception of Group 2, which had 9 subjects.
Within each study group, 8 subjects were randomly assigned to
receive SEQ ID NO.: 2 and 2 subjects were randomly assigned to
receive placebo (In Group 2 only, 1 subject received placebo). All
subjects in the study were judged by the investigator to be
healthy, normal volunteers. The test products were Glu.sup.22,25,
Leu.sup.23,28,31, Aib.sup.29, Lys.sup.26,30]hPTHrP(1-34)NH.sub.2
(SEQ ID NO.: 2) (0.1 mg/vial) and Placebo (0.9% sodium chloride
injection, USP). All subjects received a single subcutaneous (SC)
dose of Glu.sup.22,25, Leu.sup.23,28,31, Aib.sup.29,
Lys.sup.26,30]hPTHrP(1-34)NH.sub.2 (SEQ ID NO.: 2) or placebo for 7
days. The dosages and number of subjects per study group and
overall are shown below in TABLE 8.
TABLE-US-00009 TABLE 8 Number of Subjects Randomized Study Days of
Total Number Group Dose Dosing of Subjects SEQ ID NO.: 2 Placebo 1
5 .mu.g 7 10 8 2 2 20 .mu.g 7 9 8 1 3 40 .mu.g 7 10 8 2 4 80 .mu.g
7 10 8 2 Total 39 32 7
Criteria for Evaluation:
[0183] Pharmacokinetics: PK sampling for plasma SEQ ID NO.: 2 on
Days 1 and 7 was performed at the following time points: Predose (0
hour), 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 24 hours postdose.
Additionally, predose samples were taken for trough drug analysis
on Days 3 to 6. The following PK parameters were computed for Days
1 and 7: AUC.sub.(0-t), AUC.sub.(0-.infin.) (Day 1 only),
AUC.sub.(0-.tau.), AUCR (Day 1 only), C.sub.max, T.sub.max,
T.sub.last, CL/F, Kel, t.sub.1/2, and AI (Day 7 only). These
parameters were calculated from the overall (24 hours) plasma
concentration-versus-time profiles by noncompartmental methods
using WinNonlin.RTM. Pro Version 5.01 and SAS.RTM. Version 8.2.
Moreover, the ln-transformed PK parameters C.sub.max,
AUC.sub.(0-t), AUC.sub.(0-.tau.), and AUC.sub.(0-.infin.) are
presented.
Pharmacodynamics
[0184] Serum PD samples (total and ionized calcium, phosphorus,
PTH[1-84], Procollagen type 1 N-propeptide [P1NP], C-telopeptide
type 1 collagen [CTX], and 1,25-dihydroxyvitamin D [vitamin D])
were obtained at the following time points:
Serum PD Samplings were Performed at the following Time Points:
Total Calcium and Phosphorus:
[0185] Predose Days: Days -30 and -2
[0186] Days 1 and 7: Predose (0 hour), 0.5, 1, 2, 3, 4, 6, 8, 12,
and 24 hours postdose
[0187] Days 3, 4, 5, 6, and 14
Ionized Calcium:
[0188] Predose (Day -30).sup.1
[0189] Days 1 and 7: Predose (0 hour), 0.5, 1, 2, 3, 4, 6, 8, 12,
and 24 hours postdose .sup.1Deleted for Groups 2, 3, and 4 per
Amendment 3 to the protocol.
PTH(1-84) and 1, 25-dihydroxyvitamin D (Vitamin D):
[0190] Days 1 and 7: Predose (0 hour), 0.5, 1, 2, 3, 4, 6, 8, 12,
and 24 hours postdose
Procollagen Type 1 N-propeptide (P1NP) and C-telopeptide Type 1
Collagen (CTX):
[0191] Predose (Day -2), Days 3, 8, and 14.
Urine PD Samples (Calcium, Phosphorus, Cyclic AMP [c-AMP], and
Creatinine) were Obtained at the Following Time Points:
[0192] Day -1/Day 1: -24 to -18 hours, -18 to -12 hours, and -12
hours to 0 hours. Days 1 and 7: 0 to 6 hours, 6 to 12 hours, and 12
to 24 hours.
[0193] The following parameters in urine were presented for each of
the PD markers using SAS.RTM. Version 8.2: Volume (Vol.),
concentration (Conc.), amount (Ae), cumulative amount (Cum. Ae),
and excretion rates.
Statistical Methods:
Pharmacokinetics:
[0194] Plasma concentration and PK parameters for SEQ ID NO.: 2
were listed by subject and summarized by treatment and day using
descriptive statistics (mean, standard deviation [SD], coefficient
of variation [CV %], standard error of the mean [SEM], sample size
[N], minimum [min], maximum [max], and median). Additionally,
geometric means and ln-transformed values were provided for
C.sub.max and AUCs.
[0195] Dose proportionality was evaluated for Days 1 and 7 SEQ ID
NO.: 2 data using the following power model:
Ln(Y)=.beta..sub.0+.beta. Ln Dose+.epsilon.
where Y represents the PK parameters AUC.sub.(0-.infin.),
AUC.sub.(0-.tau.), AUC.sub.(0-t), and C.sub.max. Dose
proportionality requires that .beta.=1 for dose-dependent
parameters.
[0196] The model was used to calculate the 95% confidence intervals
(CI) for the slope of the ln-transformed PK parameters AUCs and
C.sub.max. Dose proportionally was concluded if the 95% CI for the
PK parameters included the value of 1.
[0197] For those cases in which dose proportionality could not be
concluded within all doses investigated, dose proportionality
analysis was performed for the first 3 (by excluding the highest
dose) and the last 3 doses (by excluding the lowest dose).
Pharmacokinetic/Pharmacodynamic Analysis:
[0198] The plots of plasma SEQ ID NO.: 2 and calcium concentrations
did not indicate a clear relationship between SEQ ID NO.: 2 and
calcium concentrations; therefore, no PK/PD modeling work was
performed for this study.
Pharmacodynamics:
[0199] The data for each PD marker in serum (total and ionized
calcium, phosphorus, PTH(1-84), 1,25-dihydroxyvitamin D, P1NP, and
CTX) and in urine (calcium, phosphorus, c-AMP, and creatinine)
following SEQ ID NO.: 2 and placebo doses were listed for each
subject and summarized by SEQ ID NO.: 2 dose using descriptive
statistics (mean, SD, CV %, SEM, N, min, max, and median).
Pharmacokinetic Results:
[0200] The arithmetic mean and the SD of plasma SEQ ID NO.: 2 PK
parameters following subcutaneous (SC) administration of SEQ ID
NO.: 2 doses for Days 1 and 7 are presented Table 9 and Table
10.
TABLE-US-00010 TABLE 9 Summary of Plasma SEQ ID NO.: 2
Pharmacokinetic Parameters Following 5 .mu.g Thro.mu.gh 80 .mu.g
SEQ ID NO.: 2 Doses-Day 1 Treatment A Treatment B Treatment C
Treatment D Pharmacokinetic Mean .+-. SD Mean .+-. SD Mean .+-. SD
Mean .+-. SD Parameters (N) (N) (N) (N) C.sub.max (pg/mL) 43.1 .+-.
10.7 115 .+-. 53.9 223 .+-. 99.0 310 .+-. 54.3 (7) (8) (8) (8)
T.sub.max (hr)# 0.566 (0.531, 1.00) 0.296 (0.250, 0.624) 0.494
(0.262, 0.579) 0.752 (0.251, 1.01) (7) (8) (8) (8) T.sub.last (hr)#
2.01 (1.50, 4.00) 2.01 (1.00, 4.00) 4.00 (1.51, 6.01) 7.00 (4.00,
12.0) (7) (8) (8) (8) AUC.sub.0-t (pg * hr/mL) 78.439 .+-. 45.472
160.52 .+-. 110.83 419.89 .+-. 275.15 949.89 .+-. 493.58 (7) (8)
(8) (8) AUC.sub.0-inf (pg * hr/mL) 187.36 .+-. 54.536 257.17 .+-.
119.05 592.94 .+-. 281.40 1055.6 .+-. 513.61 (4) (5) (6) (8)
AUC.sub.0-tau (pg * hr/mL) 186.92 .+-. 54.397 257.16 .+-. 119.02
592.90 .+-. 281.37 1053.2 .+-. 511.27 (4) (5) (6) (8) t.sub.1/2
(hr) 2.59 .+-. 0.690 1.05 .+-. 0.314 1.65 .+-. 0.254 2.30 .+-.
0.715 (4) (5) (6) (8) K.sub.el (1/hr) 0.282 .+-. 0.0722 0.713 .+-.
0.229 0.428 .+-. 0.0603 0.335 .+-. 0.127 (4) (5) (6) (8) AUCR 0.521
.+-. 0.111 0.828 .+-. 0.0449 0.838 .+-. 0.0703 0.892 .+-. 0.0369
(4) (5) (6) (8) CL/F (L/hr) 28.56 .+-. 8.727 94.20 .+-. 46.04 84.15
.+-. 46.04 94.61 .+-. 51.09 (4) (5) (6) (8) ln (C.sub.max) 3.741
.+-. 0.2153 4.643 .+-. 0.4890 5.331 .+-. 0.4130 5.722 .+-. 0.1732
(7) (8) (8) (8) ln (AUC.sub.0-t) 4.241 .+-. 0.5166 4.821 .+-.
0.8221 5.844 .+-. 0.6783 6.740 .+-. 0.5206 (7) (8) (8) (8) ln
(AUC.sub.0-inf) 5.200 .+-. 0.3016 5.456 .+-. 0.4957 6.280 .+-.
0.5203 6.855 .+-. 0.5063 (4) (5) (6) (8) ln (AUC.sub.0-tau) 5.198
.+-. 0.3007 5.456 .+-. 0.4956 6.280 .+-. 0.5202 6.853 .+-. 0.5050
(4) (5) (6) (8) #= Tmax and Tlast are presented as Median (Minimum,
Maximum) Treatment A = Administration of a Single SC Dose of 5
.mu.g SEQ ID NO.: 2 for Seven Days Treatment B = Administration of
a Single SC Dose of 20 .mu.g SEQ ID NO.: 2 for Seven Days Treatment
C = Administration of a Single SC Dose of 40 .mu.g SEQ ID NO.: 2
for seven Days Treatment D = Administration of a Single SC Dose of
80 .mu.g SEQ ID NO.: 2 for Seven Days
TABLE-US-00011 TABLE 10 Summary of Plasma SEQ ID NO.: 2
Pharmacokinetic Parameters Following 5 .mu.g Thro.mu.gh 80 .mu.g
SEQ ID NO.: 2 Doses-Day 7 Treatment A Treatment B Treatment C
Treatment D Pharmacokinetic Mean .+-. SD Mean .+-. SD Mean .+-. SD
Mean .+-. SD Parameters (N) (N) (N) (N) C.sub.max (pg/mL) 40.8 .+-.
7.63 109 .+-. 19.2 207 .+-. 77.7 436 .+-. 68.8 (6) (8) (8) (8)
T.sub.max (hr)# 1.05 (0.514, 1.53) 0.512 (0.250, 3.05) 0.492
(0.349, 1.00) 0.507 (0.500, 1.00) (6) (8) (8) (8) T.sub.last (hr)#
2.53 (1.50, 4.08) 3.00 (1.11, 4.00) 3.49 (2.00, 8.02) 6.00 (4.00,
8.02) (6) (8) (8) (8) AUC.sub.0-t (pg * hr/mL) 80.704 .+-. 30.441
171.58 .+-. 82.031 407.98 .+-. 219.70 1003.0 .+-. 383.45 (6) (8)
(8) (8) AUC.sub.0-tau (pg * hr/mL) . .+-. . 228.20 .+-. 95.154
481.88 .+-. 226.19 1080.3 .+-. 408.57 (0) (6) (8) (8) t.sub.1/2
(hr) . .+-. . 1.05 .+-. 0.244 1.43 .+-. 0.397 1.69 .+-. 0.425 (0)
(6) (8) (8) K.sub.el (1/hr) . .+-. . 0.694 .+-. 0.165 0.527 .+-.
0.192 0.437 .+-. 0.124 (0) (6) (8) (8) CL/F (L/hr) . .+-. . 103.9
.+-. 53.01 102.0 .+-. 53.34 82.74 .+-. 26.95 (0) (6) (8) (8) AI .
.+-. . 1.10 .+-. 0.369 0.844 .+-. 0.0673 1.12 .+-. 0.353 (0) (5)
(6) (8) ln (C.sub.max) 3.694 .+-. 0.1912 4.682 .+-. 0.1835 5.266
.+-. 0.4068 6.065 .+-. 0.1628 (6) (8) (8) (8) ln (AUC.sub.0-t)
4.325 .+-. 0.4085 5.039 .+-. 0.5086 5.888 .+-. 0.5352 6.851 .+-.
0.3623 (6) (8) (8) (8) ln (AUC.sub.0-tau) . .+-. . 5.351 .+-.
0.4532 6.078 .+-. 0.4879 6.927 .+-. 0.3581 (0) (6) (8) (8) #= Tmax
and Tlast are presented as Median (Minimum, Maximum) Treatment A =
Administration of a Single SC Dose of 5 .mu.g SEQ ID NO.: 2 for
Seven Days Treatment B = Administration of a Single SC Dose of 20
.mu.g SEQ ID NO.: 2 for Seven Days Treatment C = Administration of
a Single SC Dose of 40 .mu.g SEQ ID NO.: 2 for Seven Days Treatment
D = Administration of a Single SC Dose of 80 .mu.g SEQ ID NO.: 2
for Seven Days
[0201] Overall, administration of increasing doses of SEQ ID NO.: 2
resulted in increasing rate and extent of exposure to SEQ ID NO.:
2. SEQ ID NO.: 2 was characterized by a rapid absorption following
SC doses as mean C.sub.max was achieved within approximately 1
hour. Moreover, SEQ ID NO.: 2 had a short half-life with mean
t.sub.1/2 ranging from 1.05 hours to 2.59 hours. Apparent clearance
was 28.56 L/hr following the lowest dose (5 .mu.g) and ranged from
82.74 L/hr to 103.9 L/hr following the 20, 40, and 80 .mu.g doses
and, with the exception of the lowest dose remained fairly stable
with increased doses of SEQ ID NO.: 2.
[0202] The results indicated that exposure to SEQ ID NO.: 2 was
relatively comparable between Days 1 and 7 following SEQ ID NO.: 2
doses. Mean AI values ranged from 0.844 to 1.12, indicating that
dr.mu.g accumulation was negligible following multiple dosing of
SEQ ID NO.: 2. Moreover, mean PK parameters values of T.sub.max,
t.sub.1/2, and CL/F were comparable between Days 1 and 7.
[0203] The dose proportionality assessment of exposure to SEQ ID
NO.: 2 in plasma resulting from SC doses of SEQ ID NO.: 2 is
presented in the following table.
TABLE-US-00012 Dose Proportionality Analysis of Plasma SEQ ID NO.:
2 Following 5 .mu.g Through 80 .mu.g SEQ ID NO.: 2 Doses
Pharmacokinetic Standard Day Parameters Slope Error 95% CI 1 Cmax
0.77861 0.1375 (0.4935, 1.0637) AUC (0-t) 0.90714 0.1237 (0.6542,
1.1600) AUC (0-inf) 0.99565 0.2024 (0.5685, 1.4228) 7 Cmax 0.99804
0.0985 (0.7938, 1.2023) AUC (0-tau) 1.14127 0.1649 (0.7974, 1.4852)
Dose proportionality was concluded if the CI for the ln-transformed
parameters included the value of 1. Dose proportionality for Cmax
and AUC (0-inf) was concluded following 20 .mu.g, 40 .mu.g, and 80
.mu.g SEQ ID NO.: 2 doses. Dose proportionality for AUC (0-t) and
AUC (0-tau) was concluded following 5 .mu.g, 20 .mu.g, 40 .mu.g,
and 80 .mu.g SEQ ID NO.: 2 doses. Parameters were ln-transformed
prior to analysis.
Pharmacodynamic Results:
[0204] Pharmacodynamic Markers in Serum:
[0205] Total calcium concentrations in serum remained within the
reference range except for two subjects (placebo) and three
subjects receiving the SEQ ID NO.: 2 doses. On Days 1 and 7,
following SC administration of 5 to 80 .mu.g SEQ ID NO.: 2 or
placebo, mean total calcium levels marginally (.+-.0.6 mg/dL)
changed from predose levels. Total serum calcium concentrations
following SEQ ID NO.: 2 doses mostly remained above the placebo
level.
[0206] While in the first two doses, the majority of the ionized
calcium measurements including baseline values were out of the
reference range, in the second 2 doses, all the measurements were
within the reference range.
[0207] The 95% CI of the slopes for ln-transformed PK parameters
C.sub.max, AUC.sub.(0-t), AUC.sub.(0-.tau.), and
AUC.sub.(0-.infin.) indicated that, within the SEQ ID NO.: 2 dose
range studied, the increases in PK parameters were
dose-proportional (95% CI included the value of 1). While dose
proportionality for C.sub.max and AUC.sub.(0-.infin.) was concluded
only following 20, 40, and 80 .mu.g SEQ ID NO.: 2 doses, dose
proportionality for AUC.sub.(0-.tau.) and AUC.sub.(0-t) was
concluded following all SEQ ID NO.: 2 doses investigated.
[0208] Mean baseline-adjusted ionized calcium levels slightly
increased up to 0.04.+-.0.02 mmol/L following 40 .mu.g SEQ ID NO.:
2 dose on Day 1 and up to 0.05.+-.0.02 mmol/L following 80 .mu.g
SEQ ID NO.: 2 dose on Day 7. Like total calcium, mean ionized
calcium levels following SEQ ID NO.: 2 doses were generally higher
than the mean values following placebo dose.
[0209] With the exception of 24 hours on Day 1, and 8 to 12 hours
on Day 7, serum phosphorus concentrations following SEQ ID NO.: 2
and placebo doses remained below the predose levels on Days 1 and
7. Also, serum phosphorus concentrations following SEQ ID NO.: 2
doses were below the placebo concentration levels on both days.
[0210] Serum PTH (1-84) concentrations following SEQ ID NO.: 2
doses remained below the predose levels and placebo dose during
most of the sampling times on both days. Serum PTH (1-84)
concentrations following placebo dose consistently stayed above the
baseline.
[0211] 1,25-dihydroxyvitamin D concentrations in serum following
SEQ ID NO.: 2 and placebo doses generally remained at predose
levels on both Days 1 and 7, except following the 40 and 80 .mu.g
SEQ ID NO.: 2 doses which steadily rose above the predose levels
after 2 hours postdose on Day 1 and most of the time on Day 7.
Serum 1,25-dihydroxyvitamin D concentrations following SEQ ID NO.:
2 doses were mostly higher than placebo levels on both days.
[0212] P1NP concentrations in serum following SEQ ID NO.: 2 and
placebo doses generally stayed near predose levels on Days 3, 8,
and 14, except for 80 .mu.g SEQ ID NO.: 2 dose which consistently
stayed above baseline (maximum increase was up to 18.+-.12 ng/mL)
including Day 14. Mean P1NP serum levels following all doses of SEQ
ID NO.: 2 showed some non-significant dose dependent elevation on
Day 8. Mean serum CTX concentrations following SEQ ID NO.: 2 and
placebo doses generally remained at or around the predose levels
except following the 20 .mu.g SEQ ID NO.: 2 dose where the
concentrations consistently stayed above predose levels. The
maximum increase at 0.15.+-.0.18 ng/mL from baseline was within 1
SD.
Pharmacodynamic Markers in Urine:
[0213] On Day 1, mean urinary excretion rates of calcium following
SEQ ID NO.: 2 doses and placebo subjects were approximately at
predose levels. On Day 7, however, while mean urinary excretion
rates of calcium following 40 and 80 .mu.g SEQ ID NO.: 2 doses
fluctuated at predose levels, those following the 5 and 20 .mu.g
SEQ ID NO.: 2 and placebo doses dropped below the predose
levels.
[0214] Mean urinary excretion rates of phosphorus following SEQ ID
NO.: 2 doses fluctuated around the predose levels. Mean phosphorus
excretion rates following SEQ ID NO.: 2 doses on the last 2
intervals of Days 1 and 7 were lower than the first interval and at
times fell below the predose levels.
[0215] Mean urinary excretion rates of c-AMP increased following
SEQ ID NO.: 2 doses on both Days 1 and 7 but sharply dropped to
predose and at times below the predose levels by the end of the
sampling intervals.
[0216] Mean urinary excretion rates of creatinine following SEQ ID
NO.: 2 and placebo doses fluctuated around the predose levels on
Days 1 and 7.
[0217] While this invention has been particularly shown and
described with references to example embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
scope of the invention encompassed by the appended claims.
Sequence CWU 1
1
2134PRTHomo Sapiens 1Ala Val Ser Glu His Gln Leu Leu His Asp Lys
Gly Lys Ser Ile Gln1 5 10 15Asp Leu Arg Arg Arg Phe Phe Leu His His
Leu Ile Ala Glu Ile His 20 25 30Thr Ala234PRTArtificial
Sequencesynthetic peptide 2Ala Val Ser Glu His Gln Leu Leu His Asp
Lys Gly Lys Ser Ile Gln1 5 10 15Asp Leu Arg Arg Arg Glu Leu Leu Glu
Lys Leu Leu Xaa Lys Leu His 20 25 30Thr Ala
* * * * *