U.S. patent application number 11/377980 was filed with the patent office on 2006-10-05 for methods for monitoring and guiding therapeutic suppression of parathyroid hormone in renal patients having secondary hyperparathyroidism.
Invention is credited to Thomas L. Cantor.
Application Number | 20060223119 11/377980 |
Document ID | / |
Family ID | 26670921 |
Filed Date | 2006-10-05 |
United States Patent
Application |
20060223119 |
Kind Code |
A1 |
Cantor; Thomas L. |
October 5, 2006 |
Methods for monitoring and guiding therapeutic suppression of
parathyroid hormone in renal patients having secondary
hyperparathyroidism
Abstract
The present invention relates to kits for monitoring and guiding
therapeutic suppression of parathyroid hormone in renal patients
having secondary hyperparathyroidism. The kit provides means to
measure the level of a PTH agonist and a PTH antagonist in a
patient, and it further provides means for administering a PTH
suppressing therapeutic to the patient so as to reduce the level of
a PTH agonist and minimize the level of parathyroid hormone
antagonist.
Inventors: |
Cantor; Thomas L.; (El
Cajon, CA) |
Correspondence
Address: |
MORRISON & FOERSTER LLP
12531 HIGH BLUFF DRIVE
SUITE 100
SAN DIEGO
CA
92130-2040
US
|
Family ID: |
26670921 |
Appl. No.: |
11/377980 |
Filed: |
March 17, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10286465 |
Nov 1, 2002 |
7056655 |
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11377980 |
Mar 17, 2006 |
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10002818 |
Nov 2, 2001 |
6524788 |
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10286465 |
Nov 1, 2002 |
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Current U.S.
Class: |
435/7.1 ;
436/501; 514/167 |
Current CPC
Class: |
G01N 2800/10 20130101;
A61K 31/59 20130101; G01N 2800/046 20130101; A61P 19/00 20180101;
A61K 33/06 20130101; G01N 33/78 20130101; G01N 2800/347 20130101;
A61P 13/12 20180101 |
Class at
Publication: |
435/007.1 ;
514/167; 436/501 |
International
Class: |
G01N 33/53 20060101
G01N033/53; A61K 31/59 20060101 A61K031/59; G01N 33/566 20060101
G01N033/566 |
Claims
1-42. (canceled)
43. A kit for monitoring and guiding therapeutic suppression of
parathyroid hormone in a renal patient having secondary
hyperparathyroidism, the kit comprising: a) means for monitoring
the level of a parathyroid hormone (PTH) agonist in the patient; b)
means for monitoring the level of a PTH antagonist in the patient;
and c) means for administering to the patient a therapeutic that
suppresses PTH agonist, whereby the amount of therapeutic
administered is adjusted such that PTH agonist levels are decreased
and the level of PTH antagonist is minimized.
44. The kit of claim 43, wherein the therapeutic is selected from
the group consisting of vitamin D or vitamin D analogue treatment,
calcium treatment, or calcimimetic administration.
45. The kit of claim 44, wherein the Vitamin D analogue comprises
paricalcitrol, calcitriol, maxacalcitol, alfacalcidol, calcifediol,
or ergocalciferol.
46. The kit of claim 43, further comprising instructions for
use.
47. The kit of claim 43, wherein the PTH agonist comprises a
contiguous segment of human PTH having the amino acid sequence SEQ
ID NO: 1, wherein said segment starts at position 1 of human PTH,
and ends at any position from position 34 through position 84 of
human PTH.
48. The kit of claim 43, wherein the PTH antagonist comprises a
contiguous portion of human PTH having the amino acid sequence SEQ
ID NO: 1, wherein said portion starts at any position from position
2 through position 33 of human PTH, ends at any position from
position 35 through position 84 of human PTH, and has a minimal
length of three amino acid residues.
49. The kit of claim 43, wherein the PTH agonist is a peptide
having an amino acid sequence of human PTH.sub.1-84 set forth in
SEQ ID NO: 1.
50. The kit of claim 43, wherein the PTH antagonist is a peptide
having an amino acid sequence of human PTH7-84 (SEQ ID NO: 7).
51. The kit of claim 43, wherein the means for measuring the PTH
antagonist level in a patient comprises means for measuring a total
PTH level in a patient, whereby the PTH antagonist level in the
patient is determined by subtracting the PTH agonist level in the
patient from the total PTH level in the patient.
52. The kit of claim 43, wherein the means for determining a PTH
antagonist level comprises an antibody that distinguishes PTH
antagonist from PTH agonist.
53. The kit of claim 43, wherein the means for determining a PTH
agonist level comprises an antibody that distinguishes PTH agonist
from PTH antagonist.
54. A kit for monitoring and guiding therapeutic suppression of
parathyroid hormone in a renal patient having secondary
hyperparathyroidism, the kit comprising: means for monitoring the
level of a PTH antagonist in the patient; and means for
administering to the patient a therapeutic that suppresses PTH
agonist, whereby the amount of therapeutic administered is adjusted
such that the level of PTH antagonist in the patient is
minimized.
55. The kit of claim 54, wherein the PTH antagonist is a peptide
having an amino acid sequence of human PTH7-84 (SEQ ID NO: 7).
56. The kit of claim 54, wherein the means for determining a PTH
antagonist level comprises an antibody that distinguishes PTH
agonist from PTH antagonist.
57. The kit of claim 54, wherein the PTH agonist is a peptide
having an amino acid sequence of human PTH.sub.1-84 set forth in
SEQ ID NO: 1.
58. The kit of claim 54, wherein the therapeutic is selected from
the group consisting of vitamin D or vitamin D analogue treatment,
calcium treatment, or calcimimetic administration.
59. The kit of claim 58, wherein the vitamin D analogue comprises
paricalcitrol, calcitriol, maxacalcitol, alfacalcidol, calcifediol,
or ergocalciferol.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application
Ser. No. 10/286,465, filed Nov. 1, 2002, now allowed, which is a
continuation in part of U.S. patent application Ser. No.
10/002,818, filed Nov. 2, 2001, now issued as U.S. Pat. No.
6,524,788, each of which is incorporated herein by reference.
TECHNICAL FIELD
[0002] The present invention relates to novel methods for
monitoring and guiding therapeutic suppression of parathyroid
hormone in renal patients having secondary hyperparathyroidism. One
determines and monitors the level of parathyroid hormone agonist
and parathyroid hormone antagonist in the renal patient. The
parathyroid hormone suppressing therapeutic is administered to the
patient so as to minimize the level of parathyroid hormone
antagonist.
BACKGROUND ART
[0003] Calcium plays an indispensable role in cell permeability,
the formation of bones and teeth, blood coagulation, transmission
of nerve impulse, and normal muscle contraction. The concentration
of calcium ions in the blood is, along with calcitrol and
calcitonin, regulated mainly by parathyroid hormone (PTH). Although
calcium intake and excretion may vary, PTH serves through a
feedback mechanism to maintain a steady concentration of calcium in
cells and surrounding fluids. When serum calcium lowers, the
parathyroid glands secrete PTH, affecting the release of stored
calcium. When serum calcium increases, stored calcium release is
retarded through lowered secretions of PTH.
[0004] The complete form of human PTH, sometimes referred to in the
art as hPTH but referred to in the present invention as an example
of PTH agonist, is a unique 84 amino acid peptide (SEQ ID NO: 1),
as is shown in FIG. 1. Researchers have found that this peptide has
an anabolic effect on bone that involves a domain for protein
kinase C activation (amino acid residues 28 to 34) as well as a
domain for adenylate cyclase activation (amino acid residues 1 to
7). However, various catabolic forms of clipped or fragmented PTH
peptides also are found in circulation, most likely formed by
intraglandular or peripheral metabolism. For example, whole PTH can
be cleaved between amino acids 34 and 35 to produce a (1-34) PTH
N-terminal fragment and a (35-84) PTH C-terminal fragment.
Likewise, clipping can occur between either amino acids 36 and 37
or 37 and 38. Recently, a large PTH fragment referred to as
"non-(1-84) PTH" has been disclosed which is clipped closer to the
N-terminal end of PTH. (See R. LePage et al., "A non-(1-84)
circulating parathyroid hormone (PTH) fragment interferes
significantly with intact PTH commercial assay measurements in
uremic samples," Clin. Chem. ( 1998 ); 44: 805-810.)
[0005] The clinical need for accurate measurement of PTH is well
demonstrated. Serum PTH level is one of the most important indices
for patients with the following diseases: familial hypocalciuria;
hypercalcemia; multiple endocrine neoplasia types I and II;
osteoporosis; Paget's bone disease; primary
hyperparathyroidism--caused by primary hyperplasia or adenoma of
the parathyroid glands; pseudohypoparathyroidism; and renal
failure, which can cause secondary hyperparathyroidism.
[0006] PTH plays a role in the course of disease in a patient with
chronic renal failure. Renal osteodystrophy (RO) is a complex
skeletal disease comprising osteitis fibrosa cystica (caused by PTH
excess), osteomalacia--unmineralized bone matrix (caused by vitamin
D deficiency), extraskeletal calcification/ossification (caused by
abnormal calcium and phosphorus metabolism), and adynamic low bone
turnover disease (contributed to by PTH suppression). Chronic renal
failure patients can develop RO. Failing kidneys increase serum
phosphorus hyperphosphoremia) and decrease 1,25-dihydroxyvitamin D
(1,25-D) production by the kidney. The former results in secondary
hyperparathyroidism from decreased gastrointestinal calcium
absorption and osteitis fibrosa cystica from increased PTH in
response to an increase in serum phosphorus. The later causes
hypocalcemia and osteomalacia. With the onset of secondary
hyperparathyroidism, the parathyroid gland becomes less responsive
to its hormonal regulators because of decreased expression of its
calcium and vitamin D receptors. Serum calcium drops. RO can lead
to digital gangrene, bone pain, bone fractures, and muscle
weakness.
[0007] For chronic renal failure patients with secondary
hyperparathyroidism, a number of different therapeutic treatments
are available. One can administer calcium carbonate so as to
directly adjust the available calcium ion level. However, with the
increasing incidence of ectopic calcification, increasing calcium
intake is often not desirable. One can administer calcimimetics,
such as AMG073 made by Amgen, Inc. of Thousand Oaks, Calif.
However, AMG073 has not been approved for use in the USA. One can
administer vitamin D analogues, (such as the Calcijex.RTM. or
Zemplar.RTM. brands made by Abbott Labs of Abbott Park, Ill.;
Rocaltrol.RTM. brand made by Roche Laboratories of Basle,
Switzerland; Oxarol brand made by Chugai Pharmaceutical), so as to
lower PTH. However, researchers have found that vitamin D analogues
can oversuppress PTH, thereby leading to adynamic low bone turnover
disease setting the patient at risk of ectopic and vascular
calcification. (See the package insert for Zemplar.RTM., Abbott
Reference 06-9998-R1-Rev, April 1998. See the package insert for
Rocaltrol.RTM., Roche Laboratories, inc. November 1998 Product
identification Guide, page 334.)
[0008] Researchers have also found that a large circulating PTH
fragment (e.g., cyclase inactive parathyroid hormone) functions as
a naturally occurring PTH antagonist. Cyclase inactive PTH has been
found to be useful, alongside whole PTH, as an indicator in
separating untreated end stage renal disease (ESRD) patients with
high bone turnover from those with adynamic low bone turnover. (See
Faugere, M. C. et alia. "Improved Assessment of Bone Turnover by
the PTH 1-84/large C-PTH fragments ratio in ESRD patients", Kidney
International 2001; 60: 1460-1468.) Moreover, researchers have
found that cyclase inactive PTH can cause adynamic low bone
turnover by inhibiting the formation of osteoclasts, bone
resorption, and bone turnover. (See Divieti, P. et alia, "In vitro
Inhibition of Bone Resorption by Human PTH (7-84)" J. Bone Miner
Res 2001 :Suppl 1, S307. See also Faugere, M. C. et alia, "The
Effects of PTH (1-84) on bone turnover are Antagonized by PTH
(7-84) in Thyroparathyroidectomized and Nephrectomized Rats"; J Am
Soc Nephrol 12:2001, 764A.)
[0009] Determining circulating biologically active PTH levels in
humans has been challenging. One major problem is that PTH is found
at low levels, normally 10 pg/mL to 65 pg/mL. Coupled with
extremely low circulating levels is the problem of the
heterogeneity of PTH and its many circulating fragments. In many
cases, immunoassays have faced substantial and significant
interference from circulating PTH fragments. For example, some
commercially available PTH kits have almost 100% cross-reactivity
with the non- (1-84) PTH fragment, (see the LePage article).
[0010] PTH immunoassays have varied over the years. One early
approach is a double antibody precipitation immunoassay found in
U.S. Pat. No. 4,369,138 to Arnold W. Lindall et alia. A first
antibody has a high affinity for a (65-84) PTH fragment. A
radioactive labeled (65-84) PTH peptide is added to the sample with
the first antibody to compete for the endogenous unlabeled peptide.
A second antibody is added which binds to any first antibody and
radioactive labeled PTH fragment complex, thereby forming a
precipitate. Both precipitate and supernatant can be measured for
radioactive activity, and endogenous PTH levels can be calculated
therefrom.
[0011] In an effort to overcome PTH fragment interference,
immunoradiometric two-site assays for intact PTH (I-PTH) have been
introduced, such as Allegro.RTM. Intact PTH assay by the Nichol's
Institute of San Juan Capistrano, Calif. In one version, a capture
antibody specifically binds to the C-terminal portion of hPTH while
a labeled antibody specifically binds to the N-terminal portion of
the captured hPTH. In another, two monoclonal antibodies were used,
both of which attached to the N-terminal portion of hPTH.
Unfortunately, these assays have problems in that they measure but
do not discriminate between whole PTH and non-whole PTH peptide
fragments. This inability comes to the fore in hyperparathyroid
patients and renal failure patients who have significant endogenous
concentrations of large, non-whole PTH fragments.
[0012] Recently, Scantibodies Laboratory, Inc. of Santee, Calif.
USA introduced a series of novel kits that allow for the accurate
measurement of both PTH agonist and PTH antagonist. The PTH agonist
assay is a direct measurement, while the PTH antagonist assay is a
calculated value from the difference of the PTH agonist assay value
and a total PTH (including both PTH agonist and PTH antagonist)
assay value. A number of unexpected advantages have become
available to the physician, including the first non-invasive method
for assisting in the differentiation of secondary hyperparathyroid
patients with HBT and ALBT.
DISCLOSURE OF THE INVENTION
[0013] In one embodiment, a method is provided for monitoring and
guiding therapeutic suppression of parathyroid hormone in renal
patients having secondary hyperparathyroidism comprising: a)
determining and monitoring the level of parathyroid hormone (PTH)
agonist in a renal patient having secondary hyperparathyroidism; b)
determining and monitoring the level of PTH antagonist in the
patient; and c) administering a therapeutic to the patient that
suppresses PTH agonist whereby the amount of therapeutic
administered is adjusted such that the level of PTH antagonist is
minimized. Frequently, the therapeutic is administered in
increasing increments from a nominal amount. In addition,
frequently steps a) and b) are performed using a sample obtained
from a renal patient, which sample may be a serum, plasma and/or
blood sample.
[0014] In one aspect the renal patient is already receiving the
therapeutic, and therapeutic administration is terminated for a
time sufficient to allow the patient to return to at least a
relatively non-suppressed state. In one aspect, this timing may be
measured with respect to parathyroid gland status and/or bone
status. Relatedly, the time sufficient to allow the patient to
return to at least a relatively non-suppressed state is frequently
between about three months to about six months, with respect to
bone status. The time sufficient to allow the patient to return to
at least a relatively non-suppressed state is frequently between
about three minutes to about twenty minutes, with respect to
parathyroid gland status. On occasion, the time sufficient to allow
the patient to return to at least a relatively non-suppressed state
is between about two weeks to about six weeks, or sometimes about
four weeks. In addition, while guiding therapy of these patients,
the amount of administered therapeutic may be adjusted until the
level of PTH antagonist is minimized.
[0015] In another aspect of the present invention the therapeutic
being administered is selected from the group consisting of
calcium, vitamin D, vitamin D analogues, and calcimimetics (e.g.,
AMG073). Frequently, when vitamin D analogs are used, they may be
selected from paricalcitrol, calcitriol, maxacalcitol,
alfacalcidol, calcifediol, or ergocalciferol.
[0016] In a further aspect of the present invention, the PTH
agonist level is compared with the PTH antagonist level. In another
aspect, the PTH agonist level is compared with the total
parathyroid hormone level. In yet another aspect, the PTH
antagonist level is compared with the total parathyroid hormone
level. In a further aspect, the PTH agonist level is compared with
the PTH antagonist level in the form of a ratio or proportion.
[0017] In one aspect, the PTH antagonist level is determined by
determining a total PTH level and determining a PTH agonist level
followed by subtracting the PTH agonist level from the total PTH
level.
[0018] In another aspect, the PTH agonist level and the PTH
antagonist level are determined using an immunoassay. In yet
another aspect, the PTH agonist level is determined using an
antibody that distinguishes PTH agonist from PTH antagonist. In a
further aspect, frequently the PTH antagonist level is determined
using an antibody that distinguishes PTH agonist from PTH
antagonist.
[0019] In one embodiment, the method above may further comprise
monitoring for vascular calcification in the patients. Frequently,
vascular calcification is monitored by monitoring alkaline
phosphatase levels in the patient.
[0020] In another embodiment, the PTH agonist levels and the PTH
antagonist levels are compared, individually or together, with
corresponding bone biopsy data. In one aspect, the ratio or
proportion of PTH agonist to PTH antagonist will be compared with
corresponding bone biopsy data. On occasion, bone biopsy data will
be used to verify the results of PTH agonist level and/or PTH
antagonist level determinations and/or PTH agonist/antagonist
ratios. Frequently, the PTH agonist level and the PTH antagonist
level determinations are compared with corresponding bone biopsy
data after administration of the therapeutic. On occasion, this
comparison after administration of the therapeutic is useful to
monitor the PTH therapy of patients. Frequently, such treatment is
directed to decreasing PTH agonist levels and minimizing PTH
antagonist levels.
[0021] In a further aspect, the PTH agonist comprises a contiguous
portion of human PTH having an amino acid sequence set forth in SEQ
ID NO:1 (PTH.sub.1-84), and the PTH agonist has the following
characteristics: the N-terminal amino acid residue of the PTH
agonist starts at position 1 of the PTH.sub.1-84; and the
C-terminal amino acid residue of the PTH agonist ends at any
position spanning position 34 through position 84 of the
PTH.sub.1-84. In another aspect, the PTH antagonist comprises a
contiguous portion of human PTH having an amino acid sequence set
forth in SEQ ID NO: 1 (PTH.sub.1-84), and the PTH antagonist has
the following characteristics: the N-terminal amino acid residue of
the PTH antagonist starts at any position spanning position 2
through position 33 of the PTH.sub.1-84; the C-terminal amino acid
residue of the PTH antagonist ends at any position spanning
position 35 through position 84 of the PTH.sub.1-84; and the PTH
antagonist has a minimal length of three amino acid residues.
[0022] In another embodiment, a method is provided for monitoring
and guiding therapeutic suppression of parathyroid hormone in renal
patients having secondary hyperparathyroidism comprising: a)
obtaining a sample from a renal patient having secondary
hyperparathyroidism; b) determining and monitoring the level of PTH
antagonist in the renal patient; and c) administering a therapeutic
to the patient capable of suppressing a PTH agonist, whereby the
amount of therapeutic administered is adjusted such that the level
of PTH antagonist is minimized in the patient.
[0023] In a further embodiment, kits are provided that are useful
for monitoring and guiding therapeutic suppression of parathyroid
hormone in renal patients having secondary hyperparathyroidism
comprising: a) means for monitoring the level of parathyroid
hormone (PTH) agonist in a renal patient having secondary
hyperparathyroidism; b) means for monitoring the level of PTH
antagonist in the patient; and c) means for administering a
therapeutic to the patient that suppresses PTH agonist whereby the
amount of therapeutic administered is adjusted such that PTH
agonist levels are decreased and the level of PTH antagonist is
minimized.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] FIG. 1 is a diagrammatic view of human wPTH.
[0025] FIG. 2 is a graph comparing PTH measurement parameters over
time for patients receiving a PTH suppressant therapy.
DETAILED DESCRIPTION OF THE INVENTION
[0026] For clarity of disclosure, and not by way of limitation, the
detailed description of the invention is divided into the
subsections that follow.
[0027] A. Definitions
[0028] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of ordinary skill in the art to which this invention belongs. All
patents, applications, published applications and other
publications referred to herein are incorporated by reference in
their entirety. If a definition set forth in this section is
contrary to or otherwise inconsistent with a definition set forth
in the patents, applications, published applications and other
publications that are herein incorporated by reference, the
definition set forth in this section prevails over the definition
that is incorporated herein by reference.
[0029] As used herein, "a" or "an" means "at least one" or "one or
more."
[0030] As used herein, "parathyroid hormone (PTH) agonist" refers
to the complete molecule of PTH or a fragment, derivative or analog
thereof that stimulates osteoclasts formation and bone turnover to
increase blood calcium levels. PTH agonist further refers to
peptides which have PTH agonist properties. Other names of PTH
include parathormone and parathyrin. For purposes herein, the name
"parathyroid hormone (PTH)" is used herein, although all other
names are contemplated. It is intended to encompass PTH agonist
with conservative amino acid substitutions that do not
substantially alter its biological activity. Suitable conservative
substitutions of amino acids are known to those of skill in this
art and may be made generally without altering the biological
activity of the resulting molecule. Those of skill in this art
recognize that, in general, single amino acid substitutions in
non-essential regions of a polypeptide do not substantially alter
biological activity (see, e.g., Watson et al., MOLECULAR BIOLOGY OF
THE GENE, 4th Edition, 1987, The Bejamin/Cummings Pub. Co., p.224).
PTH agonist assay values may be obtained by measuring a sample with
a Scantibodies Whole PTH Assay or a Scantibodies CAP Assay or a
3.sup.rd generation PTH Assay or a Nichols BioIntact PTH assay or
an Immutopics Human Bioactive PTH assay. "Cyclase activating PTH,"
"whole PTH," and "CAP" are representative examples of PTH
agonists.
[0031] As used herein, "parathyroid hormone (PTH) antagonist"
refers to a PTH fragment or derivative that counters the effect of
a PTH agonist or otherwise lacks PTH agonist activity. It is
intended to encompass PTH antagonist with conservative amino acid
substitutions that do not substantially alter its activity.
Suitable conservative substitutions of amino acids are known to
those of skill in this art and may be made generally without
altering the biological activity of the resulting molecule. Those
of skill in this art recognize that, in general, single amino acid
substitutions in non-essential regions of a polypeptide do not
substantially alter biological activity (see, e.g., Watson, et al.
MOLECULAR BIOLOGY OF THE GENE, 4th Edition, 1987, The
Bejamin/Cummings Pub. co., p.224). "Cyclase inactive PTH" and "CIP"
are representative examples of PTH antagonists.
[0032] As used herein, the terms "total PTH," "intact PTH" and
"total intact PTH" are interchangeable and refer to an assay
directed at measuring PTH agonist and PTH antagonist levels.
[0033] As used herein, a "functional derivative or fragment" of PTH
agonist or PTH antagonist refers to a derivative or fragment of PTH
that still substantially retains its function as a PTH agonist or
PTH antagonist. Normally, the derivative or fragment retains at
least 50% of its PTH agonist or PTH antagonist activity.
Preferably, the derivative or fragment retains at least 60%, 70%,
80%, 90%, 95%, 99% and 100% of its PTH agonist or PTH antagonist
activity. It is also possible that a functional derivative or
fragment of PTH agonist or PTH antagonist has higher PTH agonist or
PTH antagonist activity than a parent molecule from which the
functional derivative or fragment is derived from.
[0034] As used herein, "treatment" means any manner in which the
symptoms of a condition, disorder or disease are ameliorated or
otherwise beneficially altered. Treatment also encompasses any
pharmaceutical use of the compositions herein.
[0035] As used herein, "disease or disorder" refers to a
pathological condition in an organism resulting from, e.g.,
infection or genetic defect, and characterized by identifiable
symptoms.
[0036] As used herein, "adynamic low bone turnover disease" refers
to a variety of disorders involving abnormal PTH agonist and/or
antagonist levels in a person. This definition is non-limiting in
that it does not refer to only one specific disease, it refers to a
variety of disorders that may result from abnormal PTH or PTH
component levels in a person. As PTH levels are tied to bone
turnover rate, abnormally low levels of PTH agonist, abnormally low
levels of PTH agonist/antagonist ratios, and abnormally high levels
of PTH antagonist may lead to abnormally low bone turnover in a
person. In a person, this type of state may indicate the presence
of, or susceptibility to, an adynamic low bone turnover disease.
Conversely, abnormally high levels of PTH agonist, abnormally high
levels of PTH agonist/antagonist ratios, and abnormally low levels
of PTH antagonist may lead to abnormally high bone turnover in a
person.
[0037] As used herein the term "sample" refers to anything which is
suspected of containing an analyte for which an analyte assay is
desired. The sample may be a biological sample, such as a
biological fluid or a biological tissue. Examples of biological
fluids include urine, blood, plasma, serum, saliva, semen, stool,
sputum, cerebral spinal fluid, tears, mucus, amniotic fluid or the
like. Biological tissues are aggregate of cells, usually of a
particular kind together with their intercellular substance that
form one of the structural materials of a human, animal, plant,
bacterial, fungal or viral structure, including connective,
epithelium, muscle and nerve tissues. Examples of biological
tissues also include organs, tumors, lymph nodes, arteries and
individual cell(s). Frequently, samples obtained for use in the
present invention contain, or are suspected of containing, levels
of PTH agonist and/or PTH antagonist that are detectable through
methods described and contemplated herein.
[0038] As used herein, the term "vitamin D analog" refers to any
available source of synthetic vitamin D from a variety of sources
for clinical or experimental use. For example, vitamin D analogs
may be obtained from sources such as Abbott Laboratories, Bone Care
International, Hoffman-La Roche, Ltd., Chugai Pharmaceutical Co.,
Amgen, Inc., NPS Pharmaceuticals, Inc, Kirin Brewing Company, Ltd.,
Sumitomo Corp., etc. Examples of vitamin D analogs available from
these sources include Calcijex.RTM., Zemplar.RTM., Hectoral.RTM.,
Rocaltrol.RTM., Oxarol, etc. In addition, the active ingredients in
a variety of synthetic vitamin D sources contemplated herein may
include paricalcitrol, calcitriol, maxacalcitol, alfacalcidol,
calcifediol, or ergocalciferol.
[0039] B. Risk and therapy determinations based on PTH agonist
levels and PTH antagonist levels
[0040] The present invention relates to novel methods for
monitoring and guiding therapeutic suppression of parathyroid
hormone in renal patients having secondary hyperparathyroidism. In
one aspect, one determines and monitors the level of PTH agonist
and PTH antagonist in the renal patient. In another aspect, one
determines and monitors the level of PTH agonist in the renal
patient. Frequently, the parathyroid hormone suppressing
therapeutic is administered to the patient so as to minimize the
level of PTH antagonist.
[0041] Secondary hyperparathyroidism is a common disease in renal
compromised patients, especially those with ESRD. Virtually all
ESRD patients have bone disease and mineral metabolism disorders,
either high bone turnover disease or adynamic low bone turnover
disease. Elevated levels of PTH agonist (with respect to PTH
antagonist) lead to high bone turnover disease (HBT). Elevated
levels of PTH antagonist (with respect to PTH agonist) lead to
adynamic low bone turnover disease (ALBT). In general, the more
medically serious of these two disorders is ALBT due to the higher
risk of soft tissue calcification that accompanies this disorder.
In ALBT ectopic tissue calcification results in vascular stenosis
(including occlusion of coronary arteries) and aortic rigidity.
Therefore, ALBT patients are subject to a higher likelihood of
serious medical complications due to a circulatory system failure,
such as myocardial infarction than those with HBT. One reason for
this is that it has been difficult in the past to find a reliable
therapeutic treatment for ALBT. In addition, the use of therapeutic
PTH suppression can lead to therapeutic PTH over-suppression which,
in turn, leads to ALBT. In other words, due to the lack of a
reliable indicator, PTH agonist suppressant therapy can
inadvertently lead to ALBT due to PTH over-suppression.
[0042] Difficulty with implementing PTH suppression therapy can be
seen in the setting of at least ten different PTH target
recommendations within the past decade. The net result of this
uncertainty in therapeutic indicators is that the incidence of
vascular calcification has been reported to be 88% for ESRD
patients. (See Goodman, W. et al., "Coronary Artery Calcification
in Young Adults with End Stage Renal Disease Who Are Undergoing
Dialysis"; NEJM 2000, May 18; 342:20, 1478-1483.)
[0043] A novel finding leading to the present invention is that
while the PTH agonist level decreases in response to the
administration of PTH agonist suppressants, PTH antagonist levels
may not. Moreover, oversuppression through the use of PTH agonist
suppressants may increase the PTH antagonist level. For an
untreated renal patient with an elevated PTH agonist level, the
administration of a PTH suppressant will also initially suppress
the PTH antagonist level. However, as the PTH suppressant dosage
increases, the level of PTH antagonist will reach a minimal level
and then start to increase before the level of PTH agonist stops
decreasing. To avoid inducing ALBT in such patients, the
administration of PTH suppressant should be adjusted so that the
PTH antagonist level reaches and stays about the minimal level
mentioned above. Ordinarily skilled artisans know that this level
may vary from patient to patient, but can determine what is best
for a particular patient through monitoring the PTH level response
to therapy.
[0044] Preferably, PTH agonist levels are measured directly using
an assay that does not detect PTH antagonist either in blood,
plasma, or serum. For example, Scantibodies Laboratory Whole PTH
Assay or Scantibodies Laboratory CAP Assay are appropriate
assays.
[0045] PTH antagonist levels should be measured using an assay that
either directly detects PTH antagonist (but not PTH agonist) or
indirectly through a total PTH measurement that measures the sum of
the PTH agonist and PTH antagonist. An indirect measurement
subtracts the PTH agonist value from the total PTH value, deriving
the PTH antagonist value. Thus, one should use a total PTH assay
that is designed to have essentially 100% cross-reactivity with PTH
antagonist and PTH agonist. For example, Scantibodies Laboratory
Total Intact PTH Assay or Scantibodies Laboratory Intact PTH Assay
are appropriate assays.
[0046] The present method should be used when a PTH suppressant
type therapy is used. Applicable treatments include calcium
administration, vitamin D and vitamin D analogue administration,
and calcimimetic administration. For untreated ESRD patients, the
therapeutic should be administered in increasing increments from an
amount nominal for the selected PTH suppressant. If the patient is
already receiving PTH suppressant type therapy, one can terminate
the therapeutic administration for a time sufficient to allow the
patient to return to a relatively non-suppressed state. The time it
takes to return to a relatively non-suppressed state may range from
about 3 to 5 minutes to about 20 minutes up to about three to about
six months.
[0047] The relatively non-suppressed state may either account for
parathyroid gland status and/or bone status. For example, without
being bound by theory, a person may return to a relatively
non-suppressed state with respect to the parathyroid gland status
in about 3 to about 5 minutes, frequently ranging up to about 20
minutes. This is because the PTH levels in a patient, with respect
to parathyroid gland status, my change within minutes of a change
in the dose of PTH suppressant, and a return to a relatively
non-suppressed state might be considered to occur within minutes.
Often times, the return to a relatively non-suppressed state, with
respect to the parathyroid gland status, may range from about 10
minutes up to one or more hours or days. Also, for example, without
being bound by theory, a person may return to a relatively
non-suppressed state with respect to the bone status in about 3
weeks to about 6 months, frequently ranging between about 3 to
about 6 months. This range may be 3 months, about 4 months, about 5
months or about 6 months. Often times, the return to a relatively
non-suppressed state may range from about 3 or about 4 weeks to
about 6 weeks. Also, frequently the time it takes to return to a
relatively non-suppressed state may be measured in days or similar
increments within the larger range of weeks or months, e.g.,
between about one to about 6 or about 7 days within a week
span.
[0048] As an alternative to discontinuing PTH type suppressant
therapy, if a patient is already receiving PTH type suppressant
therapy, one can adjust the amount of therapeutic, up or down,
until the level of PTH antagonist is minimized. Frequently, one
would take the PTH agonist level into account while adjusting
therapeutic to minimize the level of PTH antagonist. The
administering physician can easily determine the appropriate dosage
on a patient-by-patient basis. The skilled artisan appreciates that
the change in dosage for PTH suppressant may differ from that of
another. However, the physician can always be guided by the present
procedure to affect the best compromise for each individual patient
between PTH over-suppression and abnormally elevated PTH.
[0049] C. Parathyroid Hormone Antagonists
[0050] In general, a PTH antagonist of the present invention
comprises a contiguous portion of human PTH having an amino acid
sequence set forth in SEQ ID NO: 1 (PTH.sub.1-84), or a nucleic
acid encoding said portion of human PTH, and said PTH antagonist
has the following characteristics: a) the N-terminal amino acid
residue of said PTH antagonist starts at any position spanning
position 2 through position 33 of said PTH.sub.1-84; b) the
C-terminal amino acid residue of said PTH antagonist ends at any
position spanning position 35 through position 84 of said
PTH.sub.1-84; and c) said PTH antagonist has a minimal length of
three amino acid residues. Preferably, the PTH antagonist is in the
form of a pharmaceutical composition, which pharmaceutical
composition comprises an effective amount of the PTH antagonist and
a pharmaceutically acceptable carrier or excipient.
[0051] The N-terminal amino acid residue of the PTH antagonist can
start at any position spanning position 2 through position 33 of
said PTH.sub.1-84. For example, the N-terminal amino acid residue
of the PTH antagonist can start at position 2 of the PTH.sub.1-84.
The C-terminal amino acid residue of said PTH antagonist can end at
any position spanning position 35 through position 84 of said
PTH.sub.1-84. For example, the C-terminal amino acid residue of the
PTH antagonist can end at position 84 of the PTH.sub.1-84.
[0052] In a specific embodiment, the PTH antagonist is a protein or
a peptide, or a nucleic acid encoding said protein or peptide,
selected from the group consisting of PTH.sub.2-84, PTH.sub.3-84,
PTH.sub.4-84, PTH.sub.5-84, PTH.sub.6-84, PTH.sub.7-84,
PTH.sub.8-84, PTH.sub.9-84, PTH.sub.10-84, PTH.sub.11-84,
PTH.sub.12-84, PTH.sub.13-84, PTH.sub.14-84, PTH.sub.15-84,
PTH.sub.16-84, PTH.sub.17-84, PTH.sub.18-84, PTH.sub.19-84,
PTH.sub.20-84, PTH.sub.21-84, PTH.sub.22-84, PTH.sub.23-84,
PTH.sub.24-84, PTH.sub.25-84, PTH.sub.26-84, PTH.sub.27-84,
PTH.sub.28-84, PTH.sub.29-84, PTH.sub.30-84, PTH.sub.31-84,
PTH.sub.32-84, and PTH.sub.33-84. In another specific embodiment,
the PTH antagonist is a protein or a peptide, or a nucleic acid
encoding said protein or peptide, selected from the group
consisting of PTH.sub.7-69, PTH.sub.7-70, PTH.sub.7-71,
PTH.sub.7-72, PTH.sub.7-73, PTH.sub.7-74, PTH.sub.7-75,
PTH.sub.7-76, PTH.sub.7-77, PTH.sub.7-78, PTH.sub.7-79,
PTH.sub.7-80, PTH.sub.7-81, PTH7-82, PTH.sub.7-83 and
PTH.sub.7-84.
[0053] The PTH antagonist can have any suitable length provided
that it maintains its antagonizing activity. For example, the PTH
antagonist can have a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,
47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82 or 83 amino acid residues.
[0054] D. Parathyroid Hormone Agonists
[0055] In general, a PTH agonist of the present invention comprises
a contiguous portion of human PTH having an amino acid sequence set
forth in SEQ ID NO:1 (PTH.sub.1-84), and the PTH agonist has the
following characteristics: a) the N-terminal amino acid residue of
the PTH agonist starts at position 1 of the PTH.sub.1-84; and b)
the C-terminal amino acid residue of the PTH agonist ends at any
position spanning position 34 through position 84 of the
PTH.sub.1-84.
[0056] Without being bound by theory, the N-terminal amino acid
residue of the PTH agonist generally starts at position 1 of said
PTH.sub.1-84. For example, the N-terminal amino acid residue of the
PTH agonist can start at position 1 of the PTH.sub.1-84. The
C-terminal amino acid residue of said PTH agonist can end at any
position spanning position 34 through position 84 of said
PTH.sub.1-84. For example, the C-terminal amino acid residue of the
PTH agonist can end at position 84 of the PTH.sub.1-84.
[0057] The PTH agonist can have any suitable length provided that
it maintains its agonizing activity. For example, the PTH agonist
can have a length of 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,
45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82 or 83 amino acid residues.
[0058] The present invention is further described by the following
examples. The examples are provided solely to illustrate the
invention by reference to specific embodiments. These
exemplifications, while illustrating certain specific aspects of
the invention, do not portray the limitations or circumscribe the
scope of the disclosed invention.
[0059] E. Kits
[0060] The invention also provides for kits for carrying out the
methods of the invention. Such kits comprise in one or more
containers a means for determining and monitoring the level of
parathyroid hormone (PTH) agonist in a renal patient having
secondary hyperparathyroidism; in one or more containers, a means
for determining and monitoring the level of PTH antagonist in the
patient alone or in combination with other agents; and a means for
administering a therapeutic to the patient that suppresses PTH
agonist whereby the amount of therapeutic administered is adjusted
such that PTH agonist levels are decreased and the level of PTH
antagonist is minimized. Examples of means for determining and
monitoring PTH agonist levels in a patient comprise a variety of
PTH assays further described herein. And, examples of means for
determining and monitoring PTH antagonist levels in a patient
comprise a variety of PTH assays further described herein.
Preferred forms may be in combination with sterile saline, dextrose
solution, or buffered solution, or other pharmaceutically
acceptable sterile fluid. Alternatively, useful therapeutic
compositions may be lyophilized or dessicated; in this instance,
the kit optionally further comprises in a container a
pharmaceutically acceptable solution, preferably sterile, to
reconstitute the complex to form a solution for injection purposes.
Exemplary pharmaceutically acceptable solutions are saline and
dextrose solution.
[0061] In one aspect, a kit of the invention further comprises a
needle or syringe as a means for administering a therapeutic to a
patient, preferably packaged in sterile form, for injecting the
composition, and/or a packaged alcohol pad. Instructions are
optionally included for administration of composition by a
physician or by the patient.
[0062] Other features and advantages of the invention will be
apparent from the following detailed description, and from the
claims.
[0063] The present invention is further described by the following
examples. The examples are provided solely to illustrate the
invention by reference to specific embodiments. These
exemplifications, while illustrating certain specific aspects of
the invention, do not portray the limitations or circumscribe the
scope of the disclosed invention.
EXAMPLES
Example 1
[0064] A clinical trial was held for ninety ESRD patients. Each
patient had been receiving vitamin D suppressant therapy in
accordance with the manufacturer's guidelines. Each patient was
removed from the therapy for a washout period of four weeks, and
this was confirmed by a rise in PTH measurements after removal of
the therapeutic. PTH maxacalcitol (made by Chugai Pharmaceutical
Corporation of Tokyo, Japan) suppressant therapy was started after
the washout at a constant administration of 5.5 .mu.g intravenously
every three days. Blood samples were obtained from each patient
after the washout (week 0), six weeks after therapy restart (week
6), and twelve weeks after therapy restart (week 12). The samples
were assayed for PTH agonist levels and PTH antagonist levels using
the PTH agonist assay and total PTH assay made by Scantibodies
Laboratory, Inc. The samples were assayed for bone specific
alkaline phosphatase using a commercially available immunoassay
from Hybritech, Inc. of San Diego, Calif.
Clinical Results
[0065] The results of the assays for the ninety patients are shown
in FIG. 2 and the following table as median values: TABLE-US-00001
TABLE Time Time Time Parameter 0 Weeks 6 weeks 12 weeks Total PTH
886 531 525 pg/ml (41% decrease) (1% decrease) PTH agonist 609 361
331 pg/ml (41% decrease) (8% decrease) PTH antagonist 276 160 194
pg/ml (42% decrease) (21% increase) Alkaline Phosphatase 396 344
290 U/ml (13% decrease) (16% decrease)
[0066] The Table shows how the PTH suppressant lowered both the PTH
agonist and the total PTH values. After six weeks, the PTH agonist
and the total PTH values have decreased by 41%. However, after
twelve weeks, these values have decreased a mere 8% (PTH agonist)
and 0.8% (total PTH). These results indicate that the PTH
suppressant is having difficulty driving the PTH agonist levels
down any further. Moreover, the levels of total PTH and PTH agonist
are still commonly regarded as being above normal levels (of less
than 37 pg/ml for PTH agonist and less than 65 pg/ml for total PTH,
see, e.g., Nussbaum S R, et al., "Highly sensitive two-site
immunoradiometric assay ofparathyrin, and its clinical utility in
evaluating patients with hypercalcemia," Clin. Chem. 1987;
33:1364-67) for non-ESRD patients.
[0067] The Table provided above indicates the difference in
response of PTH antagonist to the PTH suppressant versus the total
PTH and PTH agonist response. After six weeks, the PTH antagonist
level has decreased by 42%, consistent with the PTH agonist levels
after six weeks. However, after twelve weeks, the PTH antagonist
level increases by 21%, in contrast to PTH agonist levels. The
continued use of PTH suppressant at this level will elevate the PTH
antagonist levels. Over a period of time, the increase in PTH
antagonist levels will lead to ALBT and subsequent vascular
calcification (as confirmed in the Table by the further drop in
alkaline phosphatase at twelve weeks). These patients need to have
the PTH suppressant dosage lowered to maintain the PTH antagonist
at a minimal (in this case 160 pg/ml) level. The medical benefit
realized in reducing the PTH agonist level beyond a particular
level, even by a small percentage, is far outweighed by the harm
caused by the resulting elevation of the PTH antagonist level. This
rise in the PTH antagonist level may cause ALBT by inhibiting
osteoclast formation, a necessary component in healthy bone
modeling involving bone resorption and bone turnover.
Example 2
[0068] To verify that PTH agonist and PTH antagonist concentrations
and the PTH agonist/antagonist ratio accurately discriminate
between high and low bone turnover in renal patients, bone biopsy
data is obtained from renal patients having secondary
hyperparathyroidism. See Faugere, M-C, et. al., Kidney Int'l. 2001;
60:1460-68. Bone biopsy data will also verify that calculation and
evaluation of both PTH agonist and PTH antagonist level data in
renal patients provides a more useful therapeutic and prognostic
indicator than evaluation of PTH agonist data alone.
[0069] Experimental Design
[0070] Patients with a total PTH greater than 200 pg/ml (as
measured by an Intact PTH assay), will have PTH agonist and PTH
antagonist levels and the PTH agonist/antagonist ratio determined
by Scantibodies.RTM. CAP PTH assay(PTH agonist), Scantibodies.RTM.
Whole PTH assay (PTH agonist), Scantibodies.RTM. total intact PTH
assay (total PTH) and/or Scantibodies.RTM. intact PTH assay (total
PTH). Those with a total PTH level greater than 400 pg/ml, or
between 200-400 pg/ml, and a PTH agonist/antagonist ratio less than
1.5 will undergo double tetracycline bone labeling and outpatient
percutaneous needle biopsy of the pelvic crest bone, under local
anesthesia.
[0071] These patients will also have their bone specific alkaline
phosphatase and TRAP (tartrate resistant acid phosphatase) measured
through known means. In addition, x-ray data will be obtained from
each patient through the use of mamographic x-ray techniques known
in the art.
[0072] Patient Population & Selection Criteria
[0073] A large cross-section of hemodialysis and peritoneal
dialysis patients will be evaluated in this study. All dialysis
patients will be included in the initial patient population
selection. However, exclusion criteria includes patients that are
generally considered too ill to participate in the study, those
whose anticoagulation cannot be discontinued for about 48 to 72
hours, and those that are unable to cooperate with study
requirements. At least 30 to 50 percent of the dialysis population
is likely to have PTH values in the range required to be considered
for bone biopsy.
[0074] Results
[0075] Bone biopsy results are correlated with results of the PTH
level assays to verify the accuracy of the PTH agonist/antagonist
ratio, versus the PTH agonist measurement alone, in determining a
patient's bone turnover rate. Biochemical and radiological data are
used to verify these results. As known clinical co-morbidities and
therapies have an effect on bone turnover, the effects of these are
taken into account in evaluating the data.
[0076] The bone biopsy data confirm the efficacy of the PTH
agonist/antagonist level comparison and therapy is guided based on
a comparison of PTH agonist levels with PTH antagonist levels.
Therapy is provided to reduce PTH agonist levels and to minimize
PTH antagonist levels in the patients.
[0077] After twelve months, biochemical and radiological parameters
are reanalyzed, and repeat bone biopsies are performed in order to
assess the clinical benefit of the PTH agonist/antagonist ratio
analysis approach.
[0078] The above examples are included for illustrative purposes
only and are not intended to limit the scope of the invention. Many
variations to those described above are possible. Since
modifications and variations to the examples described above will
be apparent to those of skill in this art, it is intended that this
invention be limited only by the scope of the appended claims.
[0079] Citation of the above publications or documents is not
intended as an admission that any of the foregoing is pertinent
prior art, nor does it constitute any admission as to the contents
or date of these publications or documents.
Sequence CWU 1
1
48 1 84 PRT Homo sapiens PEPTIDE (1)...(84) human parathyroid
hormone peptide fragment 1 Ser Val Ser Glu Ile Gln Leu Met His Asn
Leu Gly Lys His Leu Asn 1 5 10 15 Ser Met Glu Arg Val Glu Trp Leu
Arg Lys Lys Leu Gln Asp Val His 20 25 30 Asn Phe Val Ala Leu Gly
Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser 35 40 45 Gln Arg Pro Arg
Lys Lys Glu Asp Asn Val Leu Val Glu Ser His Glu 50 55 60 Lys Ser
Leu Gly Glu Ala Asp Lys Ala Asp Val Asn Val Leu Thr Lys 65 70 75 80
Ala Lys Ser Gln 2 83 PRT Homo sapiens PEPTIDE (1)...(83) human
parathyroid hormone peptide fragment 2 Val Ser Glu Ile Gln Leu Met
His Asn Leu Gly Lys His Leu Asn Ser 1 5 10 15 Met Glu Arg Val Glu
Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn 20 25 30 Phe Val Ala
Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln 35 40 45 Arg
Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu Ser His Glu Lys 50 55
60 Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn Val Leu Thr Lys Ala
65 70 75 80 Lys Ser Gln 3 82 PRT Homo sapiens PEPTIDE (1)...(82)
human parathyroid hormone peptide fragment 3 Ser Glu Ile Gln Leu
Met His Asn Leu Gly Lys His Leu Asn Ser Met 1 5 10 15 Glu Arg Val
Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 20 25 30 Val
Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg 35 40
45 Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser
50 55 60 Leu Gly Glu Ala Asp Lys Ala Asp Val Asn Val Leu Thr Lys
Ala Lys 65 70 75 80 Ser Gln 4 81 PRT Homo sapiens PEPTIDE
(1)...(81) human parathyroid hormone peptide fragment 4 Glu Ile Gln
Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu 1 5 10 15 Arg
Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val 20 25
30 Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro
35 40 45 Arg Lys Lys Glu Asp Asn Val Leu Val Glu Ser His Glu Lys
Ser Leu 50 55 60 Gly Glu Ala Asp Lys Ala Asp Val Asn Val Leu Thr
Lys Ala Lys Ser 65 70 75 80 Gln 5 80 PRT Homo sapiens PEPTIDE
(1)...(80) human parathyroid hormone peptide fragment 5 Ile Gln Leu
Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg 1 5 10 15 Val
Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala 20 25
30 Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg
35 40 45 Lys Lys Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser
Leu Gly 50 55 60 Glu Ala Asp Lys Ala Asp Val Asn Val Leu Thr Lys
Ala Lys Ser Gln 65 70 75 80 6 79 PRT Homo sapiens PEPTIDE
(1)...(79) human parathyroid hormone peptide fragment 6 Gln Leu Met
His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val 1 5 10 15 Glu
Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu 20 25
30 Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys
35 40 45 Lys Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu
Gly Glu 50 55 60 Ala Asp Lys Ala Asp Val Asn Val Leu Thr Lys Ala
Lys Ser Gln 65 70 75 7 78 PRT Homo sapiens PEPTIDE (1)...(78) human
parathyroid hormone peptide fragment 7 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp Lys Ala Asp Val Asn Val Leu Thr Lys Ala Lys Ser Gln 65 70 75
8 77 PRT Homo sapiens PEPTIDE (1)...(77) human parathyroid hormone
peptide fragment 8 Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu
Arg Val Glu Trp 1 5 10 15 Leu Arg Lys Lys Leu Gln Asp Val His Asn
Phe Val Ala Leu Gly Ala 20 25 30 Pro Leu Ala Pro Arg Asp Ala Gly
Ser Gln Arg Pro Arg Lys Lys Glu 35 40 45 Asp Asn Val Leu Val Glu
Ser His Glu Lys Ser Leu Gly Glu Ala Asp 50 55 60 Lys Ala Asp Val
Asn Val Leu Thr Lys Ala Lys Ser Gln 65 70 75 9 76 PRT Homo sapiens
PEPTIDE (1)...(76) human parathyroid hormone peptide fragment 9 His
Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu Trp Leu 1 5 10
15 Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu Gly Ala Pro
20 25 30 Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys
Glu Asp 35 40 45 Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly
Glu Ala Asp Lys 50 55 60 Ala Asp Val Asn Val Leu Thr Lys Ala Lys
Ser Gln 65 70 75 10 75 PRT Homo sapiens PEPTIDE (1)...(75) human
parathyroid hormone peptide fragment 10 Asn Leu Gly Lys His Leu Asn
Ser Met Glu Arg Val Glu Trp Leu Arg 1 5 10 15 Lys Lys Leu Gln Asp
Val His Asn Phe Val Ala Leu Gly Ala Pro Leu 20 25 30 Ala Pro Arg
Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn 35 40 45 Val
Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala 50 55
60 Asp Val Asn Val Leu Thr Lys Ala Lys Ser Gln 65 70 75 11 74 PRT
Homo sapiens PEPTIDE (1)...(74) human parathyroid hormone peptide
fragment 11 Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu Trp Leu
Arg Lys 1 5 10 15 Lys Leu Gln Asp Val His Asn Phe Val Ala Leu Gly
Ala Pro Leu Ala 20 25 30 Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg
Lys Lys Glu Asp Asn Val 35 40 45 Leu Val Glu Ser His Glu Lys Ser
Leu Gly Glu Ala Asp Lys Ala Asp 50 55 60 Val Asn Val Leu Thr Lys
Ala Lys Ser Gln 65 70 12 73 PRT Homo sapiens PEPTIDE (1)...(73)
human parathyroid hormone peptide fragment 12 Gly Lys His Leu Asn
Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys 1 5 10 15 Leu Gln Asp
Val His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro 20 25 30 Arg
Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn Val Leu 35 40
45 Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val
50 55 60 Asn Val Leu Thr Lys Ala Lys Ser Gln 65 70 13 72 PRT Homo
sapiens PEPTIDE (1)...(72) human parathyroid hormone peptide
fragment 13 Lys His Leu Asn Ser Met Glu Arg Val Glu Trp Leu Arg Lys
Lys Leu 1 5 10 15 Gln Asp Val His Asn Phe Val Ala Leu Gly Ala Pro
Leu Ala Pro Arg 20 25 30 Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys
Glu Asp Asn Val Leu Val 35 40 45 Glu Ser His Glu Lys Ser Leu Gly
Glu Ala Asp Lys Ala Asp Val Asn 50 55 60 Val Leu Thr Lys Ala Lys
Ser Gln 65 70 14 71 PRT Homo sapiens PEPTIDE (1)...(71) human
parathyroid hormone peptide fragment 14 His Leu Asn Ser Met Glu Arg
Val Glu Trp Leu Arg Lys Lys Leu Gln 1 5 10 15 Asp Val His Asn Phe
Val Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp 20 25 30 Ala Gly Ser
Gln Arg Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu 35 40 45 Ser
His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn Val 50 55
60 Leu Thr Lys Ala Lys Ser Gln 65 70 15 70 PRT Homo sapiens PEPTIDE
(1)...(70) human parathyroid hormone peptide fragment 15 Leu Asn
Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp 1 5 10 15
Val His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala 20
25 30 Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu
Ser 35 40 45 His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val
Asn Val Leu 50 55 60 Thr Lys Ala Lys Ser Gln 65 70 16 69 PRT Homo
sapiens PEPTIDE (1)...(69) human parathyroid hormone peptide
fragment 16 Asn Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln
Asp Val 1 5 10 15 His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro
Arg Asp Ala Gly 20 25 30 Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn
Val Leu Val Glu Ser His 35 40 45 Glu Lys Ser Leu Gly Glu Ala Asp
Lys Ala Asp Val Asn Val Leu Thr 50 55 60 Lys Ala Lys Ser Gln 65 17
68 PRT Homo sapiens PEPTIDE (1)...(68) human parathyroid hormone
peptide fragment 17 Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu
Gln Asp Val His 1 5 10 15 Asn Phe Val Ala Leu Gly Ala Pro Leu Ala
Pro Arg Asp Ala Gly Ser 20 25 30 Gln Arg Pro Arg Lys Lys Glu Asp
Asn Val Leu Val Glu Ser His Glu 35 40 45 Lys Ser Leu Gly Glu Ala
Asp Lys Ala Asp Val Asn Val Leu Thr Lys 50 55 60 Ala Lys Ser Gln 65
18 67 PRT Homo sapiens PEPTIDE (1)...(67) human parathyroid hormone
peptide fragment 18 Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln
Asp Val His Asn 1 5 10 15 Phe Val Ala Leu Gly Ala Pro Leu Ala Pro
Arg Asp Ala Gly Ser Gln 20 25 30 Arg Pro Arg Lys Lys Glu Asp Asn
Val Leu Val Glu Ser His Glu Lys 35 40 45 Ser Leu Gly Glu Ala Asp
Lys Ala Asp Val Asn Val Leu Thr Lys Ala 50 55 60 Lys Ser Gln 65 19
66 PRT Homo sapiens PEPTIDE (1)...(66) human parathyroid hormone
peptide fragment 19 Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp
Val His Asn Phe 1 5 10 15 Val Ala Leu Gly Ala Pro Leu Ala Pro Arg
Asp Ala Gly Ser Gln Arg 20 25 30 Pro Arg Lys Lys Glu Asp Asn Val
Leu Val Glu Ser His Glu Lys Ser 35 40 45 Leu Gly Glu Ala Asp Lys
Ala Asp Val Asn Val Leu Thr Lys Ala Lys 50 55 60 Ser Gln 65 20 65
PRT Homo sapiens PEPTIDE (1)...(65) human parathyroid hormone
peptide fragment 20 Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val
His Asn Phe Val 1 5 10 15 Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp
Ala Gly Ser Gln Arg Pro 20 25 30 Arg Lys Lys Glu Asp Asn Val Leu
Val Glu Ser His Glu Lys Ser Leu 35 40 45 Gly Glu Ala Asp Lys Ala
Asp Val Asn Val Leu Thr Lys Ala Lys Ser 50 55 60 Gln 65 21 64 PRT
Homo sapiens PEPTIDE (1)...(64) human parathyroid hormone peptide
fragment 21 Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe
Val Ala 1 5 10 15 Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser
Gln Arg Pro Arg 20 25 30 Lys Lys Glu Asp Asn Val Leu Val Glu Ser
His Glu Lys Ser Leu Gly 35 40 45 Glu Ala Asp Lys Ala Asp Val Asn
Val Leu Thr Lys Ala Lys Ser Gln 50 55 60 22 63 PRT Homo sapiens
PEPTIDE (1)...(63) human parathyroid hormone peptide fragment 22
Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu 1 5
10 15 Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg
Lys 20 25 30 Lys Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser
Leu Gly Glu 35 40 45 Ala Asp Lys Ala Asp Val Asn Val Leu Thr Lys
Ala Lys Ser Gln 50 55 60 23 62 PRT Homo sapiens PEPTIDE (1)...(62)
human parathyroid hormone peptide fragment 23 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 1 5 10 15 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 20 25 30 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 35 40
45 Asp Lys Ala Asp Val Asn Val Leu Thr Lys Ala Lys Ser Gln 50 55 60
24 61 PRT Homo sapiens PEPTIDE (1)...(61) human parathyroid hormone
peptide fragment 24 Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val
Ala Leu Gly Ala 1 5 10 15 Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln
Arg Pro Arg Lys Lys Glu 20 25 30 Asp Asn Val Leu Val Glu Ser His
Glu Lys Ser Leu Gly Glu Ala Asp 35 40 45 Lys Ala Asp Val Asn Val
Leu Thr Lys Ala Lys Ser Gln 50 55 60 25 60 PRT Homo sapiens PEPTIDE
(1)...(60) human parathyroid hormone peptide fragment 25 Arg Lys
Lys Leu Gln Asp Val His Asn Phe Val Ala Leu Gly Ala Pro 1 5 10 15
Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp 20
25 30 Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala Asp
Lys 35 40 45 Ala Asp Val Asn Val Leu Thr Lys Ala Lys Ser Gln 50 55
60 26 59 PRT Homo sapiens PEPTIDE (1)...(59) human parathyroid
hormone peptide fragment 26 Lys Lys Leu Gln Asp Val His Asn Phe Val
Ala Leu Gly Ala Pro Leu 1 5 10 15 Ala Pro Arg Asp Ala Gly Ser Gln
Arg Pro Arg Lys Lys Glu Asp Asn 20 25 30 Val Leu Val Glu Ser His
Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala 35 40 45 Asp Val Asn Val
Leu Thr Lys Ala Lys Ser Gln 50 55 27 58 PRT Homo sapiens PEPTIDE
(1)...(58) human parathyroid hormone peptide fragment 27 Lys Leu
Gln Asp Val His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala 1 5 10 15
Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn Val 20
25 30 Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala
Asp 35 40 45 Val Asn Val Leu Thr Lys Ala Lys Ser Gln 50 55 28 57
PRT Homo sapiens PEPTIDE (1)...(57) human parathyroid hormone
peptide fragment 28 Leu Gln Asp Val His Asn Phe Val Ala Leu Gly Ala
Pro Leu Ala Pro 1 5 10 15 Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys
Lys Glu Asp Asn Val Leu 20 25 30 Val Glu Ser His Glu Lys Ser Leu
Gly Glu Ala Asp Lys Ala Asp Val 35 40 45 Asn Val Leu Thr Lys Ala
Lys Ser Gln 50 55 29 56 PRT Homo sapiens PEPTIDE (1)...(56) human
parathyroid hormone peptide fragment 29 Gln Asp Val His Asn Phe Val
Ala Leu Gly Ala Pro Leu Ala Pro Arg 1 5 10 15 Asp Ala Gly Ser Gln
Arg Pro Arg Lys Lys Glu Asp Asn Val Leu Val 20 25 30 Glu Ser His
Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn 35 40 45 Val
Leu Thr Lys Ala Lys Ser Gln 50 55 30 55 PRT Homo sapiens PEPTIDE
(1)...(55) human parathyroid hormone peptide fragment 30 Asp Val
His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp 1 5 10 15
Ala Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu 20
25 30 Ser His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn
Val 35 40 45 Leu Thr Lys Ala Lys Ser Gln 50 55 31 54 PRT Homo
sapiens PEPTIDE (1)...(54) human parathyroid hormone peptide
fragment 31 Val His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro Arg
Asp Ala 1 5 10 15 Gly Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn Val
Leu Val Glu Ser 20
25 30 His Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn Val
Leu 35 40 45 Thr Lys Ala Lys Ser Gln 50 32 53 PRT Homo sapiens
PEPTIDE (1)...(53) human parathyroid hormone peptide fragment 32
His Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly 1 5
10 15 Ser Gln Arg Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu Ser
His 20 25 30 Glu Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn
Val Leu Thr 35 40 45 Lys Ala Lys Ser Gln 50 33 52 PRT Homo sapiens
PEPTIDE (1)...(52) human parathyroid hormone peptide fragment 33
Asn Phe Val Ala Leu Gly Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser 1 5
10 15 Gln Arg Pro Arg Lys Lys Glu Asp Asn Val Leu Val Glu Ser His
Glu 20 25 30 Lys Ser Leu Gly Glu Ala Asp Lys Ala Asp Val Asn Val
Leu Thr Lys 35 40 45 Ala Lys Ser Gln 50 34 63 PRT Homo sapiens
PEPTIDE (1)...(63) human parathyroid hormone peptide fragment 34
Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 1 5
10 15 Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu
Gly 20 25 30 Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro
Arg Lys Lys 35 40 45 Glu Asp Asn Val Leu Val Glu Ser His Glu Lys
Ser Leu Gly Glu 50 55 60 35 64 PRT Homo sapiens PEPTIDE (1)...(64)
human parathyroid hormone peptide fragment 35 Leu Met His Asn Leu
Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg
Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala
Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40
45 Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala
50 55 60 36 65 PRT Homo sapiens PEPTIDE (1)...(65) human
parathyroid hormone peptide fragment 36 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp 65 37 66 PRT Homo sapiens PEPTIDE (1)...(66) human
parathyroid hormone peptide fragment 37 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp Lys 65 38 67 PRT Homo sapiens PEPTIDE (1)...(67) human
parathyroid hormone peptide fragment 38 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp Lys Ala 65 39 68 PRT Homo sapiens PEPTIDE (1)...(68) human
parathyroid hormone peptide fragment 39 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp Lys Ala Asp 65 40 69 PRT Homo sapiens PEPTIDE (1)...(69)
human parathyroid hormone peptide fragment 40 Leu Met His Asn Leu
Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg
Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala
Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40
45 Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala
50 55 60 Asp Lys Ala Asp Val 65 41 70 PRT Homo sapiens PEPTIDE
(1)...(70) human parathyroid hormone peptide fragment 41 Leu Met
His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15
Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20
25 30 Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys
Lys 35 40 45 Glu Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu
Gly Glu Ala 50 55 60 Asp Lys Ala Asp Val Asn 65 70 42 71 PRT Homo
sapiens PEPTIDE (1)...(71) human parathyroid hormone peptide
fragment 42 Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg
Val Glu 1 5 10 15 Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe
Val Ala Leu Gly 20 25 30 Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser
Gln Arg Pro Arg Lys Lys 35 40 45 Glu Asp Asn Val Leu Val Glu Ser
His Glu Lys Ser Leu Gly Glu Ala 50 55 60 Asp Lys Ala Asp Val Asn
Val 65 70 43 72 PRT Homo sapiens PEPTIDE (1)...(72) human
parathyroid hormone peptide fragment 43 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp Lys Ala Asp Val Asn Val Leu 65 70 44 73 PRT Homo sapiens
PEPTIDE (1)...(73) human parathyroid hormone peptide fragment 44
Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 1 5
10 15 Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu
Gly 20 25 30 Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro
Arg Lys Lys 35 40 45 Glu Asp Asn Val Leu Val Glu Ser His Glu Lys
Ser Leu Gly Glu Ala 50 55 60 Asp Lys Ala Asp Val Asn Val Leu Thr 65
70 45 74 PRT Homo sapiens PEPTIDE (1)...(74) human parathyroid
hormone peptide fragment 45 Leu Met His Asn Leu Gly Lys His Leu Asn
Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys Leu Gln Asp
Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu Ala Pro Arg
Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu Asp Asn Val
Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55 60 Asp Lys
Ala Asp Val Asn Val Leu Thr Lys 65 70 46 75 PRT Homo sapiens
PEPTIDE (1)...(75) human parathyroid hormone peptide fragment 46
Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 1 5
10 15 Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe Val Ala Leu
Gly 20 25 30 Ala Pro Leu Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro
Arg Lys Lys 35 40 45 Glu Asp Asn Val Leu Val Glu Ser His Glu Lys
Ser Leu Gly Glu Ala 50 55 60 Asp Lys Ala Asp Val Asn Val Leu Thr
Lys Ala 65 70 75 47 76 PRT Homo sapiens PEPTIDE (1)...(76) human
parathyroid hormone peptide fragment 47 Leu Met His Asn Leu Gly Lys
His Leu Asn Ser Met Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys
Leu Gln Asp Val His Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu
Ala Pro Arg Asp Ala Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu
Asp Asn Val Leu Val Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55
60 Asp Lys Ala Asp Val Asn Val Leu Thr Lys Ala Lys 65 70 75 48 77
PRT Homo sapiens PEPTIDE (1)...(77) human parathyroid hormone
peptide fragment 48 Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met
Glu Arg Val Glu 1 5 10 15 Trp Leu Arg Lys Lys Leu Gln Asp Val His
Asn Phe Val Ala Leu Gly 20 25 30 Ala Pro Leu Ala Pro Arg Asp Ala
Gly Ser Gln Arg Pro Arg Lys Lys 35 40 45 Glu Asp Asn Val Leu Val
Glu Ser His Glu Lys Ser Leu Gly Glu Ala 50 55 60 Asp Lys Ala Asp
Val Asn Val Leu Thr Lys Ala Lys Ser 65 70 75
* * * * *