RU2777357C2 - Dosage mode of pth compound of controlled release - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: present invention relates to a method for the treatment, control, deceleration or prevention of a condition that can be treated, controlled, decelerated or prevented, using PTH. In this case, a pharmaceutical composition contains at least one PTH compound of controlled release or its pharmaceutically acceptable salt, hydrate or solvate for use for the treatment, control, deceleration or prevention of a condition that can be treated, controlled, decelerated or prevented, using PTH. The specified pharmaceutical composition is administered of no more than once every 24 hours, with a dose of the PTH compound of controlled release that corresponds to no more than 70% molar equivalent PTH 1-84 dose administered every 24 hours, necessary for maintenance of a calcium level within normal levels for the specified 24 hour period in humans.

EFFECT: obtaining a PTH compound of controlled release.

9 cl, 20 ex

Description

The present invention relates to a pharmaceutical composition containing at least one controlled release PTH compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the treatment, control, retardation or prevention of a condition that can be treated, controlled, retarded or prevented by PTH wherein said pharmaceutical composition is administered no more frequently than once every 24 hours with a controlled release PTH compound dose that corresponds to no more than 70% of the molar equivalent dose of PTH 1-84 administered every 24 hours required to maintain normal calcium levels. levels over said 24 hour period in humans, and to methods of treating, controlling, slowing or preventing said conditions.

Hypoparathyroidism is a rare endocrine disorder of calcium and phosphate metabolism that most often results from injury or removal of the parathyroid gland during thyroid surgery. Hypoparathyroidism differs from endocrine disorders in that, until recently, it was not treated by replacement of the missing hormone, parathyroid hormone, or PTH. Conventional therapy for hypoparathyroidism includes large doses of vitamin D and oral calcium supplements, which, although often effective, are associated with marked fluctuations in blood Ca ²⁺ leading to hypercalcemia and hypocalcemia, excess urinary calcium excretion, nephrocalcinosis, and ectopic calcification affecting the vascular, basal ganglia and lens of the eye.

Calcium is the most abundant mineral in the human body and its tight regulation is essential for many important biological functions such as bone mineralization, muscle contraction, nerve conduction, hormone release and blood clotting. It is particularly important to keep the calcium concentration as stable as possible due to the high sensitivity of various cellular systems or organs, including the central nervous system, muscles and exo/endocrine glands, to small changes in Ca ²⁺ . PTH is the main regulator of calcium homeostasis.

The inappropriately low PTH relative to serum Ca ²⁺ concentration, characteristic of hypoparathyroidism, results in reduced renal tubular reabsorption of Ca ²⁺ and concomitantly increased renal tubular reabsorption of phosphate. Thus, the main biochemical disorders of hypoparathyroidism are hypocalcemia and hyperphosphatemia. Clinical signs of the disease include symptoms of hypocalcemia such as perioral numbness, paresthesia, and wrist/foot muscle spasms. Throat spasm, tetany, and seizures are serious and potentially life-threatening complications. Hyperphosphatemia and elevated calcium phosphate product contribute to the ectopic deposition of insoluble calcium phosphate complexes in soft tissues, including the vasculature, brain, kidneys, and other organs.

The standard therapy for hypoparathyroidism is oral calcium and vitamin D supplementation. The goals of therapy are to: a) improve the symptoms of hypocalcemia; b) maintaining fasting serum calcium levels within or slightly below normal; c) maintaining fasting serum phosphorus within the normal range or only slightly elevated; d) avoid or minimize hypercalciuria; e) maintain the content of calcium phosphate product at a level well below the upper limit of normal; f) avoid ectopic calcification of the kidneys (stones and nephrocalcinosis) and other soft tissues.

Several problems arise with long-term use of calcium and active vitamin D at high doses, especially with regard to hypercalciuria, kidney stones, nephrocalcinosis, and ectopic soft tissue calcification. In addition, traditional therapy with calcium and active vitamin D does not improve quality of life complaints or eliminate the bone remodeling abnormalities that are characteristic of this disease. Briefly, there is a high need for improved treatments for hypoparathyroidism.

In 2015, Natpara, PTH(1-84) was approved for once daily subcutaneous injection as a supplement to vitamin D and calcium in patients with hypoparathyroidism. Natpara, PTH (1-84), was approved for the control of hypocalcemia based on a pivotal study demonstrating that 42 percent of participants taking PTH (1-84) achieved normal blood calcium levels with reduced doses of calcium supplements and active forms of vitamin D compared to 3 percent of participants who received a placebo. After a period of time during which post-injection serum calcium levels were monitored, 71 percent of patients treated with PTH (1–84) developed hypercalcemia on one or more measurements within a 24-hour period. PTH(1–84) reduced urinary calcium excretion 2–8 hours post-injection, but urinary calcium excretion did not change over a 24-hour period. Similarly, urinary phosphate excretion only increased during the first 8 hours after PTH injection (1–84).

Although this represents an important advance in the treatment of this disease, Natpara has not been shown to reduce the incidence of hypercalcemia (increased serum calcium), hypocalcemia (low serum calcium), or hypercalciuria (increased urinary calcium) compared to traditional therapy in treated patients.

Thus, there is a high need for improved PTH-based treatments for hypoparathyroidism.

PTH (1-34), or teriparatide, was approved by the FDA in 2002 for the treatment of osteoporosis. Although not approved for this indication, PTH(1-34) has historically been used to treat hypoparathyroidism in patients who received injections twice or thrice daily. To promote more physiological levels of PTH, clinical studies have been conducted with PTH(1-34) administered via a pump pack system compared to twice-daily injections. After 6 months, delivery via a pump packaging system provided normal, stable calcium levels with minimal fluctuations and prevented the increase in serum and urinary calcium that occurs shortly after PTH injection. The marked decrease in urinary calcium excretion when PTH(1–34) is administered via a pump-packing system may indicate that PTH must be continuously exposed to the renal tubules for the effect of calcium storage in the kidneys. Delivery using the PTH (1–34) pump packing system allowed for simultaneous normalization of bone remodeling markers, serum calcium, and urinary calcium excretion. These results were achieved with a 65 percent lower daily dose of PTH (1-34) and a reduced requirement for magnesium supplementation compared to a twice-daily PTH (1-34) injection regimen.

However, continuous pump therapy is inconvenient and difficult for patients, and it is an object of the present invention to provide a more convenient therapeutic option for providing continuous exposure to PTH.

Long-term daily administration of PTH is associated with progressive loss of cortical bone due to increased bone turnover. During a 6-year follow-up of patients treated with PTH (1-84) (Rubin, JCEM 2016), markers of bone remodeling remained higher than pre-treatment values, peaking in the first years after initiation of PTH (1-84) and declining after this, but remaining significantly higher than baseline values by year 6. Bone mineral density (BMD) as determined by double X-ray absorptiometry (DXA) was consistent with the known site-specific effects of PTH, namely, an increase in the lumbar spine in the distal 1/3 radius. The observed decrease in the 1/3 radius is consistent with the known effects of intermittent PTH to increase cortical porosity and intraosseous resorption.

The object of the present invention was to provide a method for intermittent administration of PTH with improved control of serum and urinary calcium, serum phosphorus, and a lower increase in bone remodeling markers than currently used PTH treatments. Preferably "periodically" means at daily intervals, or alternatively or more preferably at weekly intervals.

In the preclinical development program of both Forteo, PTH (1-34) and Natpara, PTH (1-84), a dose-dependent increase in the incidence of osteosarcoma was observed in rats given daily injections of the PTH compound. In the Natpara study, dosing of high-dose rats was discontinued due to excessive mortality in this group, mainly from metastatic osteosarcoma. This is thought to be due to the sensitivity of rats to the anabolic effects of intermittent PTH. In contrast, long-term exposure to PTH is known to lack significant bone anabolic activity. Thus, it is an object of the present invention to provide intermittent PTH replacement therapy that provides an infusion-like PTH profile resulting in improved symptom control at a lower administered dose. Preferably "periodically" means at daily intervals, or alternatively or more preferably at weekly intervals.

Thus, there is a need for a more convenient and safer treatment of hypoparathyroidism with reduced side effects.

Therefore, the object of the present invention is to at least partially overcome the disadvantages described above.

This object is achieved by a pharmaceutical composition comprising at least one controlled release PTH compound, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the treatment, control, retardation, or prevention of a condition that can be treated, controlled, retarded, or prevented by PTH, wherein said pharmaceutical composition is administered no more frequently than once every 24 hours with a controlled release PTH compound dose that corresponds to no more than 70% of the molar equivalent dose of PTH 1-84 administered every 24 hours required to maintain calcium levels within normal levels over a specified 24-hour period in humans.

Surprisingly, this controlled release PTH compound has been found to be more effective than PTH 1-84, so fewer molar equivalents need to be administered per dose to the patient in order to achieve beneficial serum calcium levels in at least 24 hours, which improves efficacy. and reduces the risk of side effects.

It is understood that PTH 1-84 is a polypeptide with the sequence of SEQ ID NO:1.

In the context of the present invention, the terms used have the following meanings.

As used in this application, the terms "not more often than once every 24 hours" and "at least once every 24 hours" are used interchangeably and mean that the time between two consecutive administrations is 24 hours or more, which means eg that there may be, for example, 24 hours, 48 hours, 72 hours, 96 hours, 120 hours, 144 hours, or one week between two successive administrations.

As used herein, the terms "within the normal range" and "within the normal range" in relation to serum calcium levels refer to the level of calcium normally found in a subject of a given species, sex, and age, expressed as a range defined by the lower limit of normal and upper limit of normal. In humans, the normal level preferably corresponds to a serum calcium level above 8.5 mg/dl (adjusted for albumin). In humans, the upper limit of normal is below 10.5 mg/dL.

As used herein, the term "serum calcium above 8.5 mg/dL" refers to albumin-adjusted calcium concentrations.

As used in this application, the term "albumin-adjusted" in relation to calcium levels means that the measured serum calcium level is corrected for calcium bound to albumin, according to the following formula:

albumin-adjusted serum calcium (mg/dl)=measured total Ca (mg/dl)+0.8 (4.0 - serum albumin [g/dl])

As used herein, the term "unregulated" in relation to calcium levels means that the measured total calcium concentration (mg/dL) is not adjusted in relation to calcium albumin binding.

The term “molar equivalent dose” refers to the dose at which a controlled release PTH compound contains the same number of PTH molecules or PTH moieties as a particular dose of PTH 1-84 contains PTH 1-84 molecules. For example, if a controlled release PTH compound contains one PTH molecule or PTH moiety per controlled release PTH compound, the molar equivalent dose is 1 controlled release PTH compound for every 1 molecule of PTH 1-84. If a controlled release PTH compound contains two PTH molecules or PTH constituents per controlled release PTH compound, the molar equivalent dose is 1 controlled release PTH compound for every 2 molecules of PTH 1-84.

As used herein, the term “controlled release PTH compound” refers to any compound, conjugate, crystal, or mixture that contains at least one PTH molecule or PTH moiety and from which at least one PTH molecule or PTH moiety is released with a half-life of at least 12 hours.

As used herein, the terms "half-life" and "half-life" refer to the time required under physiological conditions (i.e., in an aqueous buffer, pH 7.4, 37°C) until half of the total PTH or constituents The PTH respectively contained in the controlled release PTH compound will not be released from said controlled release PTH compound.

As used herein, the term “PTH” refers to all PTH polypeptides, preferably mammalian species, more preferably human and mammalian species, more preferably human and murine species, as well as variants, analogs, orthologs, homologues and derivatives and fragments thereof, which characterized by an increase in serum calcium and renal excretion of phosphorus, and a decrease in serum phosphorus and excretion of calcium by the kidneys. The term "PTH" also refers to all PTH-related polypeptides (PTHrP), such as the polypeptide according to SEQ ID NO:121, which binds to and activates the common PTH/PTHrP1 receptor. Preferably, the term “PTH” refers to a PTH polypeptide according to SEQ ID NO:51 as well as variants, homologues and derivatives thereof that exhibit substantially the same biological activity, i. increasing serum calcium and phosphorus excretion by the kidneys, and decreasing serum phosphorus and calcium excretion by the kidneys.

Preferably, the term "PTH" refers to the following polypeptide sequences:

SEQ ID NO:1 (PTH 1-84)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAKSQ

SEQ ID NO:2 (PTH 1-83)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAKS

SEQ ID NO:3 (PTH 1-82)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAK

SEQ ID NO:4 (PTH 1-81)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKA

SEQ ID NO:5 (PTH 1-80)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTK

SEQ ID NO:6 (PTH 1-79)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLT

SEQ ID NO:7 (PTH 1-78)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVL

SEQ ID NO:8 (PTH 1-77)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNV

SEQ ID NO:9 (PTH 1-76)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVN

SEQ ID NO:10 (PTH 1-75)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADV

SEQ ID NO:11 (PTH 1-74)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKAD

SEQ ID NO:12 (PTH 1-73)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKA

SEQ ID NO:13 (PTH 1-72)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADK

SEQ ID NO:14 (PTH 1-71)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEAD

SEQ ID NO:15 (PTH 1-70)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEA

SEQ ID NO:16 (PTH 1-69)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGE

SEQ ID NO:17 (PTH 1-68)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLG

SEQ ID NO:18 (PTH 1-67)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSL

SEQ ID NO:19 (PTH 1-66)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKS

SEQ ID NO:20 (PTH 1-65)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEK

SEQ ID NO:21 (PTH 1-64)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHE

SEQ ID NO:22 (PTH 1-63)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESH

SEQ ID NO:23 (PTH 1-62)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVES

SEQ ID NO:24 (PTH 1-61)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVE

SEQ ID NO:25 (PTH 1-60)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLV

SEQ ID NO:26 (PTH 1-59)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVL

SEQ ID NO:27 (PTH 1-58)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNV

SEQ ID NO:28 (PTH 1-57)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDN

SEQ ID NO:29 (PTH 1-56)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKED

SEQ ID NO:30 (PTH 1-55)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKE

SEQ ID NO:31 (PTH 1-54)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKK

SEQ ID NO:32 (PTH 1-53)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRK

SEQ ID NO:33 (PTH 1-52)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPR

SEQ ID NO:34 (PTH 1-51)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRP

SEQ ID NO:35 (PTH 1-50)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQR

SEQ ID NO:36 (PTH 1-49)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQ

SEQ ID NO:37 (PTH 1-48)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGS

SEQ ID NO:38 (PTH 1-47)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAG

SEQ ID NO:39 (PTH 1-46)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDA

SEQ ID NO:40 (PTH 1-45)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRD

SEQ ID NO:41 (PTH 1-44)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPR

SEQ ID NO:42 (PTH 1-43)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAP

SEQ ID NO:43 (PTH 1-42)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLA

SEQ ID NO:44 (PTH 1-41)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPL

SEQ ID NO:45 (PTH 1-40)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAP

SEQ ID NO:46 (PTH 1-39)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGA

SEQ ID NO:47 (PTH 1-38)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALG

SEQ ID NO:48 (PTH 1-37)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVAL

SEQ ID NO:49 (PTH 1-36)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVA

SEQ ID NO:50 (PTH 1-35)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFV

SEQ ID NO:51 (PTH 1-34)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF

SEQ ID NO:52 (PTH 1-33)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHN

SEQ ID NO:53 (PTH 1-32)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVH

SEQ ID NO:54 (PTH 1-31)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDV

SEQ ID NO:55 (PTH 1-30)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQD

SEQ ID NO:56 (PTH 1-29)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQ

SEQ ID NO:57 (PTH 1-28)

SVSEIQLMHNLGKHLNSMERVEWLRKKL

SEQ ID NO:58 (PTH 1-27)

SVSEIQLMHNLGKHLNSMERVEWLRKK

SEQ ID NO:59 (PTH 1-26)

SVSEIQLMHNLGKHLNSMERVEWLRK

SEQ ID NO:60 (PTH 1-25)

SVSEIQLMHNLGKHLNSMERVEWLR

SEQ ID NO:61 (amidated PTH 1-84)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAKSQ; where the C-terminus is amidated

SEQ ID NO:62 (amidated PTH 1-83)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAKS; where the C-terminus is amidated

SEQ ID NO:63 (amidated PTH 1-82)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKAK; where the C-terminus is amidated

SEQ ID NO:64 (amidated PTH 1-81)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTKA; where the C-terminus is amidated

SEQ ID NO:65 (amidated PTH 1-80)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLTK; where the C-terminus is amidated

SEQ ID NO:66 (amidated PTH 1-79)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVLT; where the C-terminus is amidated

SEQ ID NO:67 (amidated PTH 1-78)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNVL; where the C-terminus is amidated

SEQ ID NO:68 (amidated PTH 1-77)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVNV; where the C-terminus is amidated

SEQ ID NO:69 (amidated PTH 1-76)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADVN; where the C-terminus is amidated

SEQ ID NO:70 (amidated PTH 1-75)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKADV; where the C-terminus is amidated

SEQ ID NO:71 (amidated PTH 1-74)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKAD; where the C-terminus is amidated

SEQ ID NO:72 (amidated PTH 1-73)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADKA; where the C-terminus is amidated

SEQ ID NO:73 (amidated PTH 1-72)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEADK; where the C-terminus is amidated

SEQ ID NO:74 (amidated PTH 1-71)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEAD; where the C-terminus is amidated

SEQ ID NO:75 (amidated PTH 1-70)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGEA; where the C-terminus is amidated

SEQ ID NO:76 (amidated PTH 1-69)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLGE; where the C-terminus is amidated

SEQ ID NO:77 (amidated PTH 1-68)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSLG; where the C-terminus is amidated

SEQ ID NO:78 (amidated PTH 1-67)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKSL; where the C-terminus is amidated

SEQ ID NO:79 (amidated PTH 1-66)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEKS; where the C-terminus is amidated

SEQ ID NO:80 (amidated PTH 1-65)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHEK; where the C-terminus is amidated

SEQ ID NO:81 (amidated PTH 1-64)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESHE; where the C-terminus is amidated

SEQ ID NO:82 (amidated PTH 1-63)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVESH; where the C-terminus is amidated

SEQ ID NO:83 (amidated PTH 1-62)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVES; where the C-terminus is amidated

SEQ ID NO:84 (amidated PTH 1-61)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLVE; where the C-terminus is amidated

SEQ ID NO:85 (amidated PTH 1-60)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVLV; where the C-terminus is amidated

SEQ ID NO:86 (amidated PTH 1-59)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNVL; where the C-terminus is amidated

SEQ ID NO:87 (amidated PTH 1-58)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDNV; where the C-terminus is amidated

SEQ ID NO:88 (amidated PTH 1-57)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKEDN; where the C-terminus is amidated

SEQ ID NO:89 (amidated PTH 1-56)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKED; where the C-terminus is amidated

SEQ ID NO:90 (amidated PTH 1-55)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKKE; where the C-terminus is amidated

SEQ ID NO:91 (amidated PTH 1-54)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRKK; where the C-terminus is amidated

SEQ ID NO:92 (amidated PTH 1-53)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPRK; where the C-terminus is amidated

SEQ ID NO:93 (amidated PTH 1-52)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRPR; where the C-terminus is amidated

SEQ ID NO:94 (amidated PTH 1-51)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQRP; where the C-terminus is amidated

SEQ ID NO:95 (amidated PTH 1-50)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQR; where the C-terminus is amidated

SEQ ID NO:96 (amidated PTH 1-49)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGSQ; where the C-terminus is amidated

SEQ ID NO:97 (amidated PTH 1-48)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAGS; where the C-terminus is amidated

SEQ ID NO:98 (amidated PTH 1-47)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDAG; where the C-terminus is amidated

SEQ ID NO:99 (amidated PTH 1-46)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRDA; where the C-terminus is amidated

SEQ ID NO:100 (amidated PTH 1-45)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPRD; where the C-terminus is amidated

SEQ ID NO:101 (amidated PTH 1-44)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAPR; where the C-terminus is amidated

SEQ ID NO:102 (amidated PTH 1-43)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLAP; where the C-terminus is amidated

SEQ ID NO:103 (amidated PTH 1-42)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPLA; where the C-terminus is amidated

SEQ ID NO:104 (amidated PTH 1-41)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAPL; where the C-terminus is amidated

SEQ ID NO:105 (amidated PTH 1-40)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGAP; where the C-terminus is amidated

SEQ ID NO:106 (amidated PTH 1-39)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALGA; where the C-terminus is amidated

SEQ ID NO:107 (amidated PTH 1-38)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVALG; where the C-terminus is amidated

SEQ ID NO:108 (amidated PTH 1-37)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVAL; where the C-terminus is amidated

SEQ ID NO:109 (amidated PTH 1-36)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFVA; where the C-terminus is amidated

SEQ ID NO:110 (amidated PTH 1-35)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNFV; where the C-terminus is amidated

SEQ ID NO:111 (amidated PTH 1-34)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF; where the C-terminus is amidated

SEQ ID NO:112 (amidated PTH 1-33)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHN; where the C-terminus is amidated

SEQ ID NO:113 (amidated PTH 1-32)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVH; where the C-terminus is amidated

SEQ ID NO:114 (amidated PTH 1-31)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQDV; where the C-terminus is amidated

SEQ ID NO:115 (amidated PTH 1-30)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQD; where the C-terminus is amidated

SEQ ID NO:116 (amidated PTH 1-29)

SVSEIQLMHNLGKHLNSMERVEWLRKKLQ; where the C-terminus is amidated

SEQ ID NO:117 (amidated PTH 1-28)

SVSEIQLMHNLGKHLNSMERVEWLRKKL; where the C-terminus is amidated

SEQ ID NO:118 (amidated PTH 1-27)

SVSEIQLMHNLGKHLNSMERVEWLRKK; where the C-terminus is amidated

SEQ ID NO:119 (amidated PTH 1-26)

SVSEIQLMHNLGKHLNSMERVEWLRK; where the C-terminus is amidated

SEQ ID NO:120 (amidated PTH 1-25)

SVSEIQLMHNLGKHLNSMERVEWLR; where the C-terminus is amidated

SEQ ID NO:121 (PTHrP)

AVSEHQLLHDKGKSIQDLRRRFFLHHLIAEIHTAEIRATSEVSPNSKPSPNTKNHPVRFGSDDEGRYLTQETNKVETYKEQPLKTPGKKKKGKPGKRKEQEKKKRRTRSAWLDSGVTGSGLEGDHLSDTSTTSLELDSRRH

More preferably, the term "PTH" refers to the sequence SEQ ID: NO 47, 48, 49, 50, 51, 52, 53, 54, 55, 107, 108, 109, 110, 111, 112, 113, 114 and 115. Even more preferably, the term "PTH" refers to the sequence SEQ ID:NO 50, 51, 52, 110, 111 and 112. In a particularly preferred embodiment of the present invention, the term "PTH" refers to the sequence SEQ ID NO:51.

As used herein, the term “PTH polypeptide variant” refers to a polypeptide from the same species that differs from a reference PTH or PTHrP polypeptide. Preferably, such a reference polypeptide is the PTH polypeptide sequence of SEQ ID NO:51. In general, differences are limited such that the amino acid sequences of the reference polypeptide and the variant are generally very similar and, in many areas, identical. Preferably, the PTH polypeptide variants are at least 70%, 80%, 90%, or 95% identical to the reference PTH or PTHrP polypeptide, preferably the PTH polypeptide of SEQ ID NO:51. By a polypeptide having an amino acid sequence at least, for example, 95% “identical” to the amino acid sequence in question, it is meant that the amino acid sequence of the polypeptide is identical to the sequence in question, except that the polypeptide sequence may include up to five amino acid changes for every 100 amino acids of the amino acid sequence in question. These substitutions in the reference sequence can occur at the amino (N-terminal) or carboxy-terminal (C-terminal) positions of the reference amino acid sequence, or at any position between these terminal positions, distributed either individually among the residues in the reference sequence, or as one or more contiguous groups in the reference sequence. The sequence in question may be either the entire amino acid sequence of the reference sequence or any fragment specified as described herein. Preferably, the sequence in question is SEQ ID NO:51.

Such PTH polypeptide variants may be naturally occurring variants, such as naturally occurring allelic variants encoded by one of several alternative forms of PTH or PTHrP occupying a given locus on a chromosome or organism, or isoforms encoded by naturally occurring splicing variants derived from one primary transcript. Alternatively, the PTH polypeptide variant may be a variant that is not known to occur naturally and that can be produced by mutagenesis techniques known in the art.

It is known in the art that one or more amino acids can be removed from the N-terminus or C-terminus of a bioactive polypeptide without significant loss of biological function. Such N- and/or C-terminal deletions are also encompassed by the term PTH polypeptide variant.

It is also known to those skilled in the art that certain amino acid sequences of PTH or PTHrP polypeptides can be changed without significantly affecting the structure or function of the polypeptide. Such mutants include deletions, insertions, inversions, repeats and substitutions chosen according to general rules known in the art to have little effect on activity. For example, guidance on how to make phenotypically silent amino acid substitutions is provided in Bowie et al. (1990), Science 247:1306-1310, which is incorporated herein by reference in its entirety, where the authors indicate that there are two main approaches to studying amino acid sequence tolerance to change.

The term PTH polypeptide also encompasses all PTH and PTHrP polypeptides encoded by PTH and PTHrP analogs, orthologs and/or species homologues. Those skilled in the art will also appreciate that PTHrP and PTHrP analogs bind to the activation of the common PTH/PTHrP1 receptor, so the term PTH polypeptide also encompasses all PTHrP analogs. As used in this application, the term “PTH analogue” refers to PTH and PTHrP o different and unrelated organisms that perform the same functions in each organism, but which do not come from the ancestral structure that the organisms' ancestors had. Instead, the similar PTH and PTHrP arose separately and then evolved to perform the same or similar functions. In other words, the analogous PTH and PTHrP polypeptides are polypeptides with completely different amino acid sequences, but which have the same biological activity, namely an increase in serum calcium and renal phosphorus excretion, and a decrease in serum phosphorus and renal calcium excretion.

As used in this application, the term “PTH ottolog” refers to PTH and PTHrP of two different species, the sequences of which are related to each other through a common homologous PTH in the ancestral species, but which have become distinct from each other during evolution.

As used herein, the term "PTH homologue" refers to PTH and PTHrP of different organisms that perform the same function in each organism and that are derived from an ancestral structure that the organisms' ancestors had in common. In other words, PTH homologous polypeptides are polypeptides with exactly similar amino acid sequences that have the same biological activity, namely an increase in serum calcium and renal phosphorus excretion, and a decrease in serum phosphorus and renal calcium excretion. Preferably, PTH polypeptide homologues can be defined as polypeptides having at least 40%, 50%, 60%, 70%, 80%, 90%, or 95% identity with a reference PTH or PTHrP polypeptide, preferably the PTH polypeptide of SEQ ID NO: 51.

Thus, a PTH polypeptide according to the present invention may be, for example: (i) a polypeptide wherein at least one of the amino acid residues has as a substituent a conservative or non-conserved amino acid residue, preferably a conservative amino acid residue, and such substituted amino acid residue can or cannot be encoded by the genetic code; and/or (ii) a polypeptide in which at least one of the amino acid residues includes a substituent group; and/or (iii) a polypeptide wherein the PTH polypeptide is fused to another compound, such as a compound to increase the half-life of the polypeptide (eg, polyethylene glycol); and/or (iv) a polypeptide in which additional amino acids are fused to a PTH polypeptide, such as a polypeptide or IgG Fc fusion region leader or secretory sequence, or a sequence that is used to purify the above form of the polypeptide, or a precursor protein sequence.

As used herein, the term “PTH polypeptide fragment” refers to any peptide containing a consecutive span of a portion of the PTH or PTHrP amino acid sequence of a polypeptide, preferably a polypeptide of SEQ ID NO:51.

More specifically, a fragment of a PTH polypeptide contains at least 6, such as at least 8, at least 10 or at least 17 consecutive amino acids of a PTH or PTHrP polypeptide, more preferably a polypeptide of the sequence SEQ ID NO:51. A PTH polypeptide fragment can be further described as a subspecies of PTH or PTHrP polypeptides containing at least 6 amino acids, where "at least 6" is defined as an integer between 6 and an integer denoting the C-terminal amino acid of a PTH or PTHrP polypeptide, preferably a polypeptide sequence SEQ ID No:51. Also included are species of PTH or PTHrP polypeptide fragments of at least 6 amino acids in length as described above, which are further specified in terms of their N-terminal and C-terminal positions. Also encompassed by the term "PTH polypeptide fragment" as separate species are all fragments of a PTH or PTHrP polypeptide of at least 6 amino acids in length as described above, which can be specifically specified by their N-terminal and C-terminal position. That is, each combination of an N-terminal and a C-terminal position that a constituent of at least 6 consecutive amino acid residues in length can occupy, on any given amino acid sequence of a PTH or PTHrP polypeptide, preferably a PTH polypeptide of sequence SEQ ID:NO51, is included in the present invention.

The term “PTH” also includes poly(amino acid) conjugates that have the sequence as described above but having a backbone containing both amide and non-amide linkages, such as ester linkages, such as depsipeptides. Depsipeptides are chains of amino acid residues in which the main chain contains both amide (peptide) and ester bonds. Accordingly, the term “side chain”, as used herein, refers either to a moiety attached to the alpha carbon of an amino acid moiety, if the amino acid moiety is connected via amine bonds, as in polypeptides, or to any moiety containing a carbon atom, attached to the backbone of a poly(amino acid) conjugate, such as in the case of depsipeptides. Preferably, the term “PTH” refers to polypeptides having a main chain formed through amide (peptide) bonds.

Since the term PTH includes the above-described variants, analogs, orthologues, homologues, derivatives, and fragments of PTH and PTHrP, all references to specific positions in the reference sequence also include the equivalent positions in the variants, analogs, orthologues, homologues, derivatives, and fragments of the PTH or PTHrP component, even unless specifically stated.

As used in this application, the term "micelle" means an aggregate of amphiphilic molecules dispersed in a liquid colloid. In aqueous solution, a typical micelle forms an aggregate with the hydrophilic portion of the surfactant molecules facing the surrounding solvent and the hydrophobic portion of the surfactant molecules facing inwards, also referred to as a "normal phase micelle". "Invers micelles" have a hydrophilic portion facing inward and a hydrophobic portion facing the surrounding solvent.

As used in this application, the term "liposome" refers to a vesicle, preferably a spherical vesicle, having at least one lipid bilayer. Preferably, the liposomes contain phospholipids, even more preferably phosphatidylcholine. The term "liposome" refers to various structures and sizes, for example, multilayer liposomal vesicles (MLV) having more than one concentric lipid bilayer with an average diameter of 100 to 1000 nm, small unilamellar liposomal vesicles (SUV) having one lipid bilayer and an average diameter of 25 to 100 nm, to large unilamellar liposomal vesicles (LUV) having one lipid bilayer and an average diameter of about 1000 µm, and giant unilamellar vesicles (GUV) having one lipid bilayer and an average diameter of 1 to 100 µm. The term "liposome" also includes elastic vesicles such as, for example, transferosomes and ethosomes.

As used in this application, the term "aquasome" refers to spherical nanoparticles with a diameter of 60 to 300 nm, which contain at least three layers of a self-organizing structure, namely a solid-state nanocrystalline core coated with an oligomeric film, on which drug molecules are adsorbed with drug modification. means or without it.

As used in this application, the term "ethosome" refers to lipid vesicles containing phospholipids and ethanol and/or isopropanol in relatively high concentration and water, having a size in the range from tens of nanometers to micrometers.

As used herein, the term “LeciPlex” refers to a positively charged phospholipid based vesicular system that contains soy PC, a cationic agent and a biocompatible solvent such as PEG 300, PEG 400, diethylene glycol monoethyl ether, polyethylene glycol tetrahydrofurfuryl alcohol ether or 2-pyrrolidone or N-methyl-2-pyrrolidone.

As used in this application, the term "niosome" refers to single-layer or multilayer vesicles containing non-ionic surfactants.

As used in this application, the term "pharmacosome" refers to ultrafine vesicular, micellar or hexagonal aggregates of lipids covalently associated with biologically active constituents.

As used herein, the term “proniosomes” refers to dry compositions of a surfactant-coated carrier that, upon rehydration and moderate agitation, produces niosomes.

As used in this application, the term "polymersome" refers to an artificial spherical vesicle containing a membrane formed from amphiphilic synthetic block copolymers, and may optionally contain an aqueous solution in its core. The polymersome has a diameter ranging from 50 nm to 5 µm or more. The term also includes syntosomes, which are polymersomes designed to contain channels that allow certain chemicals to pass through the membrane into or out of the vesicle.

As used in this application, the term "sphingosome" refers to a concentric bilayer vesicle in which the water volume is completely surrounded by a membrane lipid bilayer, mainly consisting of natural or synthetic sphingolipid.

As used in this application, the term "transferosome" refers to ultra-flexible lipid vesicles containing an aqueous core that is formed from a mixture of conventional polar and suitable lipids, activated at the edges, which facilitate the formation of highly curved bilayers, which make the transferosome highly deformable.

As used in this application, the term "ufasome" refers to a vesicle containing unsaturated fatty acids.

As used in this application, the term "pelipept" refers to a peptide containing up to and including 50 amino acid monomers.

As used in this application, the term "protein" refers to a peptide of more than 50 amino acid residues. Preferably the protein contains at most 20,000 amino acid residues, such as at most 15,000 amino acid residues, such as at most 10,000 amino acid residues, such as at most 5,000 amino acid residues, such as at most 4,000 amino acid residues, such as at most 3,000 amino acid residues, such as for example at most 2000 amino acid residues, such as at most 1000 amino acid residues.

As used in this application, the term "physiological states" refers to an aqueous buffer at pH 7.4, 37°C.

As used in this application, the term "pharmaceutical composition" refers to a composition containing one or more active ingredients, such as, for example, at least one controlled release PTH compound, and one or more excipients, as well as any product that, directly or indirectly, results from the combination, complexation, or aggregation of any two or more ingredients of the composition, or the dissociation of one or more ingredients, or other types of reactions or interactions of one or more ingredients. Accordingly, a pharmaceutical composition for use according to the present invention encompasses any composition obtained by mixing one or more controlled release PTH compounds and a pharmaceutically acceptable excipient.

As used in this application, the term "liquid composition" refers to a mixture containing a water-soluble PTH compound and one or more solvents, such as water.

The term “suspension composition” refers to a mixture containing a water-insoluble PTH compound and one or more solvents, such as water.

As used in this application, the term "dry composition" means that the pharmaceutical composition is provided in dry form. Suitable drying methods are spray drying and lyophilization, i. e. freeze drying. Such a dry prodrug composition has a residual water content of at most 10%, preferably less than 5% and more preferably less than 2%, as determined by the Karl Fischer method. Preferably, the pharmaceutical composition for use according to the present invention is dried by lyophilization.

The term “drug”, as used herein, refers to a substance, such as PTH, used to treat, alleviate, prevent, or diagnose a disease, or otherwise improve physical or mental health. When a drug is conjugated to another moiety, the moiety of the resulting product that is derived from the drug is referred to as the “bioactive moiety”.

As used herein, the term “prodrug” refers to a conjugate containing a biologically active moiety reversibly and covalently linked to a specialized protecting group via a reversible linker moiety, also referred to as a “reversible prodrug linker moiety”, which contains a reversible bond to the biologically active moiety, and where the specialized protecting group alters or reduces undesirable properties in the parent molecule. It also includes enhancing desirable drug properties and suppressing undesirable properties. A specialized non-toxic protecting group is referred to as a "carrier". The prodrug releases the reversibly and covalently bound biologically active moiety in the form of its corresponding drug. In other words, a prodrug is a conjugate containing a biologically active moiety that is reversibly and covalently conjugated to a carrier moiety via a reversible linker moiety of the prodrug, wherein the covalent or reversible conjugation of the carrier to the reversible linker moiety of the prodrug is either direct or via a spacer. Such a conjugate releases the previously conjugated biologically active moiety in the form of a free, unmodified drug.

The terms “biodegradable bond” or “reversible bond” means a bond that is hydrolytically degradable, ie. degradable, in the absence of enzymes under physiological conditions (aqueous buffer at pH 7.4, 37°C) with a half-life in the range of one hour to three months, preferably one hour to two months, even more preferably one hour to one month, even more preferably one hour to three weeks, most preferably one hour to two weeks. Accordingly, a stable bond is one having a half-life under physiological conditions (aqueous buffer at pH 7.4, 37°C) of more than three months.

As used herein, the term "traceless prodrug linker" means a reversible prodrug linker, ie. a linker moiety that reversibly and covalently links the biologically active moiety to the carrier, which, upon cleavage, releases the drug in its free form. As used in this application, the term "free form" of a drug means a drug in its unmodified, pharmacologically active form.

As used herein, the term "excipient" refers to a diluent, adjuvant, or carrier with which a therapeutic agent, such as a drug or prodrug, is administered. Such a pharmaceutical excipient may be a sterile liquid such as water and oils, including liquids of petroleum, animal, vegetable, or synthetic origin, including, but not limited to, peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is the preferred excipient when the pharmaceutical composition is administered orally. Saline and aqueous dextrose are the preferred vehicles when the pharmaceutical composition is administered intravenously. Salt solutions and aqueous solutions of dextrose and glycerol are preferably used as liquid excipients for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, milk powder, glycerin, propylene glycol, water, ethanol and the like. The composition may also contain, if desired, small amounts of wetting or emulsifying agents, pH buffering agents such as, for example, acetate, succinate, tris, carbonate, phosphate, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, MES (2 -(N-morpholine)ethanesulfonic acid) or may contain detergents such as Tween, poloxamers, poloxamines, CHAPS, Igepal, or amino acids such as, for example, glycine, lysine or histidine Such pharmaceutical compositions may take the form of solutions, suspensions, emulsions tablets, pills, capsules, powders, sustained release formulations, etc. The pharmaceutical composition may be in the form of a suppository, with conventional binders and excipients such as triglycerides.The oral formulation may contain standard excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Such compositions contain a therapeutic and an effective amount of the drug or biologically active moiety, together with a suitable amount of excipient, such that a form suitable for administration to a patient is obtained. The composition must be appropriate for the route of administration.

As used in this application, the term "reagent" means a chemical compound that contains at least one functional group for reaction with a functional group of another chemical compound or drug. It is understood that a drug containing a functional group (such as a primary or secondary amine or hydroxyl functional group) is also a reagent.

As used in this application, the term "constituent" means a part of a molecule in which one or more atoms are missing compared to the corresponding reactants. If, for example, a reagent of the formula “H-X-H” reacts with another reagent and becomes part of the reaction product, the corresponding component of the reaction product has the structure “H–X–” or “-–X–”, where each “-–” denotes attachment to another component . Accordingly, the biologically active moiety is released from the prodrug as a drug.

It is understood that if a sequence or chemical structure of a group of atoms is provided, where the group of atoms attaches to two constituents or interrupts a constituent, said sequence or structure can be attached to two constituents in any orientation, unless otherwise indicated. For example, the component "-C(O)N(R ¹ )-" can be attached to two components or interrupt the component either as "-C(O)N(R ¹ )-" or as "-N(R ¹ ) C(O)-“. Likewise, the component

, либо как

.may be attached to two components or may interrupt a component either as

, either as

.

As used in this application, the term "functional group" means a group of atoms that can react with other groups of atoms. Functional groups include, but are not limited to, the following groups: carboxylic acid (–(C=O)OH), primary and secondary amine (–NH ₂ , –NH–), maleimide, thiol (-SH), sulfonic acid ( –(O=S=O)OH), carbonate, carbamate (–O(C=O)N<), hydroxyl (–OH), aldehyde (–(C=O)H), ketone (–(C=O )–), hydrazine (>NN<), isocyanate, isothiocyanate, phosphoric acid (–O(P=O)OHOH), phosphonic acid (–O(P=O)OHH), haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine , disulfide, sulfonamides, sulfuric acid, vinyl sulfone, vinyl ketone, diazoalkane, oxirane and aziridine.

In case the controlled release PTH compound for use according to the present invention contains one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically or toxicologically acceptable salts, in particular their pharmaceutically acceptable salts. Thus, controlled release PTH compounds for use according to the present invention containing acidic groups can be used according to the present invention, for example, in the form of alkali metal salts, alkaline earth metal salts or ammonium salts. More specific examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts or ammonium salts, or with organic amines such as, for example, ethylamine, ethanolamine, triethanolamine or amino acids. A PTH controlled release compound for use according to the present invention containing one or more basic groups, i. e. groups that can be protonated can be present and used according to the present invention in the form of their addition salts with inorganic or organic acids. Examples of suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid , pivaloic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and other known acids, qualified professionals in this field. Those skilled in the art are aware of converting a basic group to a cation, such as alkylating an amine group to give a positively charged ammonium group and the corresponding salt counterion. If the controlled release PTH compound for use according to the present invention simultaneously contains acidic and basic groups, the present invention also includes, in addition to the mentioned salt forms, internal salts or betaines (zwitterions). The corresponding salts may be prepared by conventional methods known to those skilled in the art, for example by contacting these compounds with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange with other salts. The present invention also includes all salts of the compounds for use according to the present invention which, due to low physiological compatibility, are not directly suitable for use in pharmaceutical products, but which can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.

The term "pharmaceutically acceptable" means a substance that does not cause harm when administered to a patient and is preferably approved by a regulatory authority such as EMEA (Europe) and/or FDA (USA) and/or any other national regulatory authority for use in animals, preferably humans .

As used in this application, the term "about" in combination with a numerical value is used to indicate a range in the interval and including the numerical value plus and minus no more than 10% of the specified numerical value, more preferably no more than 8% of the specified value, even more preferably not more than 5% of the specified value and most preferably not more than 2% of the specified value. For example, the phrase “about 200” is used to refer to a range between and including 200+/- 10%, i.e. in the range and including 180 - 220; preferably 200+/- 8% i.e. in the interval and including 184 - 216; even more preferably in the range and including 200+/-5%, ie. in the range and including 190 - 210; and most preferably 200+/- 2%, i. e. in the range and including 196 - 204. It is clear that the percentage given as "about 20%" does not mean "20% +/- 10%", i.e. in the range and including 10 - 30%, but "about 20%" means in the range and including 18 - 22%, i.e. plus and minus 10% of the numerical value, which is 20.

As used in this application, the term "polymer" means a molecule containing repeating structural units, i. monomers linked by chemical bonds in a linear, circular, branched, crosslinked, or dendrimeric manner, or a combination thereof, which may be of synthetic or biological origin, or a combination of both. It is understood that the polymer may also contain one or more other chemical groups and/or constituents, such as, for example, one or more functional groups. Preferably, the soluble polymer has a molecular weight of at least 0.5 kDa, for example, a molecular weight of at least 1 kDa, a molecular weight of at least 2 kDa, a molecular weight of at least 3 kDa, or a molecular weight equal to at least 5 kDa. If the polymer is soluble, it preferably has a molecular weight of at most 1000 kDa, such as at most 750 kDa, such as at most 500 kDa, such as at most 300 kDa, such as at most 200 kDa, such as at most 100 kDa. It is understood that for insoluble polymers, such as hydrogels, there are no significant molecular weight ranges. It is also understood that a protein is a polymer in which amino acids are repetitive structural units, even though the side chains of each amino acid may be different.

As used in this application, the term "polymeric" means a reagent or component containing one or more polymers or polymeric components. The polymeric reactant or constituent may optionally also contain one or more other constituents, which are preferably selected from the group consisting of:

• C _1-50 alkyl, C _2-50 alkenyl, C _2-50 alkynyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl ; and

• links selected from the group containing

where

dotted lines denote attachment to the remainder of the moiety or reactant, and

-R and R ^a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

One skilled in the art will appreciate that the polymerization products obtained from a polymerization reaction do not all have the same molecular weight, but rather have a molecular weight distribution. Therefore, the molecular weight ranges, molecular weights, ranges of the amount of monomers in the polymer and the amount of monomers in the polymer, as used in this application, refer to the number average molecular weight and the average number of monomers, i.e. to the arithmetic mean of the molecular weight of the polymer or polymer fragment and the arithmetic mean of the number of monomers of the polymer or polymer fragment.

Accordingly, in a polymeric component containing "x" monomer units, any integer given for "x" therefore corresponds to the arithmetic average of the number of monomers. Any range of integers given for "x" provides a range of integers in which lies the arithmetic average of the number of monomers. An integer for "x" given as "about x" means that the arithmetic averages of the monomers lie in the range of integers x+/- 10%, preferably x+/- 8%, more preferably x+/- 5% and most preferably x+ /- 2%.

As used in this application, the term "number average molecular weight" means the usual arithmetic average of the molecular weights of individual polymers.

As used herein, the term “water-soluble” with reference to a carrier means that when such a carrier is part of a controlled release PTH compound for use according to the present invention, at least 1 g of a controlled release PTH compound containing such a water-soluble carrier, can be dissolved in one liter of water at 20°C to form a homogeneous solution. Accordingly, the term "water-insoluble" when referring to a carrier means that when such a carrier is part of a PTH of a controlled release compound for use according to the present invention, less than 1 g of a PTH of a controlled release compound containing such a water-soluble carrier can be dissolved in one liter water at 20°C to form a homogeneous solution.

As used herein, the term "water-soluble" when referring to a controlled release PTH compound means that at least 1 g of the controlled release PTH compound can be dissolved in one liter of water at 20° C. to form a homogeneous solution. Accordingly, the term "water-insoluble" when referring to a controlled release PTH compound means that less than 1 g of the controlled release PTH compound can be dissolved in one liter of water at 20° C. to form a homogeneous solution.

As used in this application, the term "hydrogel" means a hydrophilic or amphiphilic polymer network, consisting of homopolymers or copolymers, which is insoluble due to the presence of covalent chemical cross-links. Cross links form the network structure and physical integrity.

As used herein, the term "thermogel" means a compound that is a low viscosity liquid or solution having a viscosity of less than 500 cps at 25° C. at a shear rate of about 0.1/s at low temperature, the low temperature being in the range from about 0°C to about 10°C, but which is a compound with a higher viscosity of less than 10,000 centipoise at 25°C with a shear rate of about 0.1/s at a higher temperature, the higher temperature being in the range of about 30°C to about 40°C, for example at a temperature of about 37°C.

As used in this application, the term "PEG-based" in relation to the component or reagent means that the specified component or reagent contains PEG. Preferably, the PEG-based component or reagent contains at least 10% (wt./wt.) PEG, such as at least 20% (wt./wt.) PEG, such as at least 30% (wt./wt.) wt.) PEG, such as at least 40% (wt./wt.) PEG, such as at least 50% (wt./wt.), such as at least 60 (wt./wt.) PEG such as at least 70% (w/w) PEG such as at least 80% (w/w) PEG such as at least 90% (w/w) PEG such as for example at least 95%. The remaining weight percentages of the PEG-based constituent or reagent are other constituents, preferably selected from the following constituents and linkages:

• C _1-50 alkyl, C _2-50 alkenyl, C _2-50 alkynyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl ; and

• links selected from the group containing

where

dotted lines denote attachment to the remainder of the moiety or reactant, and

-R and R ^a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

As used herein, the term "PEG-based containing at least X% PEG" in relation to a constituent or reactant means that said constituent or reactant contains at least X% (w/w) units of ethylene glycol (-CH ₂ CH ₂ O–), where the ethylene glycol units may be arranged in a block, alternating or random arrangement in a constituent or reactant, and preferably all ethylene glycol units of said constituent or reactant are present in one block; the remaining weight percentages of the PEG-based constituent or reagent are other constituents, preferably selected from the following constituents and relationships:

• C _1-50 alkyl, C _2-50 alkenyl, C _2-50 alkynyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl and tetralinyl ; and

• links selected from the group containing

where

dotted lines denote attachment to the remainder of the moiety or reactant, and

-R and R ^a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

The term “based on hyaluronic acid containing at least X% hyaluronic acid” is used appropriately.

The term "substituted", as used in this application, means that one or more hydrogen atoms of a molecule or constituent is replaced by another atom or group of atoms, which are referred to as "Deputy".

Preferably, one or more additional optional substituents are independently selected from the group consisting of halogen, -CN, -COOR ^x1 , -OR ^x1 , -C(O)R ^x1 , -C(O)N(R ^x1 R ^x1a ), -S(O) ₂ N(R ^x1 R ^x1a ), -S(O)N(R ^x1 R ^x1a ), -S(O) ₂ R ^x1 , -S(O)R ^x1 , -N(R ^x1 )S(O) ₂ N(R ^x1a R ^x1b ), -SR ^x1 , -N(R ^x1 R ^x1a ), -NO ₂ , -OC(O)R ^x1 , -N(R ^x1 )C(O) R ^x1a , -N(R ^x1 )S(O) ₂ R ^x1a , -N(R ^x1 )S(O)R ^x1a , -N(R ^x1 )C(O)OR ^x1a , -N(R ^x1 )C (O)N(R ^x1a R ^x1b ), -OC(O)N(R ^x1 R ^x1a ), -T ⁰ , C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where -T ⁰ , C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally substituted with one or more -R ^x2 that are the same or different, and where C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T ⁰ -, -C(O)O-, -O-, -C(O)-, -C(O)N( R ^x3 )-, -S(O) ₂ N(R ^x3 )-, -S(O)N(R ^x3 )-, -S(O) ₂ -, -S(O)-, -N(R ^x3 )S(O) ₂ N(R ^x3a )-, -S-, -N(R ^x3 )-, -OC(OR ^x3 )(R ^x3a )-, -N(R ^x3 )C(O)N(R ^x3a )- and -OC(O)N(R ^x3 )-;

-R ^x1 , -R ^x1a , -R ^{x1b are} independently selected from the group consisting of -H, -T ⁰ , C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where -T ⁰ , C _1-50 alkyl, C _2-50 alkenyl, and C _2-50 alkynyl are optionally substituted with one or more substituents -R ^x2 that are the same or different, and where C _1-50 alkyl, C _{2- 50} alkenyl and C _2-50 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T ⁰ -, -C(O)O-, -O-, -C(O)-, -C(O) N(R ^x3 )-, -S(O) ₂ N(R ^x3 )-, -S(O)N(R ^x3 )-; -S(O) ₂ -, -S(O)-, -N(R ^x3 )S(O) ₂ N(R ^x3a )-, -S-, -N(R ^x3 )-, -OC(OR ^x3 )(R ^x3a )-, -N(R ^x3 )C(O)N(R ^x3a )- and -OC(O)N(R ^x3 )-;

each T ⁰ is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl; where each T ⁰ is independently optionally substituted with one or more substituents -R ^x2 that are the same or different;

each -R ^x2 is independently selected from the group consisting of halogen, -CN, oxo (=O), -COOR ^x4 , -OR ^x4 , -C(O)R ^x4 , -C(O)N(R ^x4 R ^x4a ) , -S(O) ₂ N(R ^x4 R ^x4a ), -S(O)N(R ^x4 R ^x4a ), -S(O) ₂ R ^x4 , -S(O)R ^x4 , -N(R ^x4 )S(O) ₂ N(R ^x4a R ^x4b ), -SR ^x4 , -N(R ^x4 R ^x4a ), -NO ₂ , -OC(O)R ^x4 , -N(R ^x4 )C(O)R ^x4a , -N(R ^x4 )S(O) ₂ R ^x4a , -N(R ^x4 )S(O)R ^x4a , -N(R ^x4 )C(O)OR ^x4a , -N(R ^x4 )C( O)N(R ^x4a R ^x4b ), -OC(O)N(R ^x4 R ^x4a ) and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogen substituents that are the same or different;

each -R ^x3 , -R ^x3a , -R ^x4 , -R ^x4a , -R ^{x4b is} independently selected from the group consisting of -H and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogen substituents that are the same or different.

More preferably, one or more additional optional substituents are independently selected from the group consisting of halogen, -CN, -COOR ^x1 , -OR ^x1 , -C(O)R ^x1 , -C(O)N(R ^x1 R ^x1a ), -S(O) ₂ N(R ^x1 R ^x1a ), -S(O)N(R ^x1 R ^x1a ), -S(O) ₂ R ^x1 , -S(O)R ^x1 , -N( R ^x1 )S(O) ₂ N(R ^x1a R ^x1b ), -SR ^x1 , -N(R ^x1 R ^x1a ), -NO ₂ , -OC(O)R ^x1 , -N(R ^x1 )C(O )R ^x1a , -N(R ^x1 )S(O) ₂ R ^x1a , -N(R ^x1 )S(O)R ^x1a , -N(R ^x1 )C(O)OR ^x1a , -N(R ^x1 ) C(O)N(R ^x1a R ^x1b ), -OC(O)N(R ^x1 R ^x1a ), -T ⁰ , C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl; where -T ⁰ , C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl are optionally substituted with one or more -R ^x2 that are the same or different, and where C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T ⁰ -, -C(O)O-, -O-, -C(O)-, -C(O)N( R ^x3 )-, -S(O) ₂ N(R ^x3 )-, -S(O)N(R ^x3 )-, -S(O) ₂ -, -S(O)-, -N(R ^x3 )S(O) ₂ N(R ^x3a )-, -S-, -N(R ^x3 )-, -OC(OR ^x3 )(R ^x3a )-, -N(R ^x3 )C(O)N(R ^x3a )- and -OC(O)N(R ^x3 )-;

each -R ^x1 , -R ^x1a , -R ^x1b , -R ^x3 , -R ^{x3a is} independently selected from the group consisting of H, halogen, C _1-6 alkyl, C _2-6 alkenyl and C _2-6 alkynyl;

each T ⁰ is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl; where each T ⁰ is independently optionally substituted with one or more -R ^x2 that are the same or different;

each -R ^x2 is independently selected from the group consisting of halogen, -CN, oxo (=O), -COOR ^x4 , -OR ^x4 , -C(O)R ^x4 , -C(O)N(R ^x4 R ^x4a ) , -S(O) ₂ N(R ^x4 R ^x4a ), -S(O)N(R ^x4 R ^x4a ), -S(O) ₂ R ^x4 , -S(O)R ^x4 , -N(R ^x4 )S(O) ₂ N(R ^x4a R ^x4b ), -SR ^x4 , -N(R ^x4 R ^x4a ), -NO ₂ , -OC(O)R ^x4 , -N(R ^x4 )C(O)R ^x4a , -N(R ^x4 )S(O) ₂ R ^x4a , -N(R ^x4 )S(O)R ^x4a , -N(R ^x4 )C(O)OR ^x4a , -N(R ^x4 )C( O)N(R ^x4a R ^x4b ), -OC(O)N(R ^x4 R ^x4a ), and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogens that are the same or different;

each -R ^x4 , -R ^x4a , -R ^{x4b is} independently selected from the group consisting of H, halogen, C _1-6 alkyl, C _2-6 alkenyl and C _2-6 alkynyl;

Even more preferably, one or more additional optional substituents are independently selected from the group consisting of halogen, -CN, -COOR ^x1 , -OR ^x1 , -C(O)R ^x1 , -C(O)N(R ^x1 R ^x1a ), -S(O) ₂ N(R ^x1 R ^x1a ), -S(O)N(R ^x1 R ^x1a ), -S(O) ₂ R ^x1 , -S(O)R ^x1 , -N (R ^x1 )S(O) ₂ N(R ^x1a R ^x1b ), -SR ^x1 , -N(R ^x1 R ^x1a ), -NO ₂ , -OC(O)R ^x1 , -N(R ^x1 )C( O)R ^x1a , -N(R ^x1 )S(O) ₂ R ^x1a , -N(R ^x1 )S(O)R ^x1a , -N(R ^x1 )C(O)OR ^x1a , -N(R ^x1 )C(O)N(R ^x1a R ^x1b ), -OC(O)N(R ^x1 R ^x1a ), -T ⁰ , C _1-6 alkyl, C _2-6 alkenyl and C _2-6 alkynyl; where -T ⁰ , C _1-6 alkyl, C _2-6 alkenyl and C _2-6 alkynyl are optionally substituted with one or more -R ^x2 that are the same or different, and where C _1-6 alkyl, C _2-6 alkenyl and C _2-6 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T ⁰ -, -C(O)O-, -O-, -C(O)-, -C(O)N( R ^x3 )-, -S(O) ₂ N(R ^x3 )-, -S(O)N(R ^x3 )-, -S(O) ₂ -, -S(O)-, -N(R ^x3 )S(O) ₂ N(R ^x3a )-,-S-, -N(R ^x3 )-, -OC(OR ^x3 )(R ^x3a )-, -N(R ^x3 )C(O)N(R ^x3a )-, and -OC(O)N(R ^x3 )-;

each -R ^x1 , -R ^x1a , -R ^x1b , -R ^x2 , -R ^x3 , -R ^{x3a is} independently selected from the group consisting of H, halogen, C _1-6 alkyl, C _2-6 alkenyl and C _{2- 6} alkynyl;

each T ⁰ is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl; where each T ⁰ is independently optionally substituted with one or more -R ^x2 that are the same or different.

Preferably, a maximum of 6-H atoms of the optionally substituted molecule are independently substituted with a substituent, e.g., 5-H atoms are independently substituted with a substituent, 4-H atoms are independently substituted with a substituent, 3-H atoms are independently substituted with a substituent, 2-H atoms are independently substituted with a substituent, or 1 -H atom is substituted with a substituent.

The term "interrupted" means that the fragment is inserted between two carbon atoms or - if the insert is at one end of the component - between a carbon atom or a heteroatom and a hydrogen atom, preferably between a carbon atom and a hydrogen atom.

As used in this application, the term "C _1-4 alkyl" alone or in combination means a straight or branched alkyl moiety having 1 to 4 carbon atoms. If present at the end of the molecule, examples of straight or branched C _1-4 alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. When two constituent molecules are linked by C _1-4 alkyl, examples of such C _1-4 alkyl groups are -CH ₂ -, -CH ₂ -CH ₂ -, -CH(CH ₃ )-, -CH ₂ -CH ₂ -CH ₂ -, -CH(C ₂ H ₅ )-, -C(CH ₃ ) ₂ -. Each C _1-4 alkyl carbon hydrogen may optionally be substituted with a substituent as defined above. Optionally, C _1-4 alkyl may be interrupted by one or more moieties as defined below.

As used in this application, the term "C _1-6 alkyl" alone or in combination means a straight or branched alkyl moiety having from 1 to 6 carbon atoms. If present at the end of the molecule, examples of straight or branched C _1-6 alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl and 3,3-dimethylpropyl. When two constituent molecules are linked by a C _1-6 alkyl group, examples of such C _1-6 alkyl groups are -CH ₂ -, -CH ₂ -CH ₂ -, -CH(CH ₃ )-, -CH ₂ -CH ₂ -CH ₂ -, -CH(C ₂ H ₅ )- and C(CH ₃ ) ₂ -. Each hydrogen atom at the C _1-6 carbon atom may optionally be substituted with a substituent as defined above. Optionally, C _1-6 alkyl may be interrupted by one or more moieties as defined below.

Accordingly, "C _1-10 alkyl", "C _1-20 alkyl" or "C _1-50 alkyl" means an alkyl chain having 1 to 10, 1 to 20, or 1 to 50 carbon atoms, respectively, where each hydrogen atom at the C _1-10 , C _1-20 or C _1-50 carbon atom may optionally be substituted with a substituent as defined above. Optionally, C _1-10 or C _1-50 alkyl may be interrupted by one or more moieties as defined below.

As used herein, the term "C _2-6 alkenyl" alone or in combination means a straight or branched hydrocarbon moiety containing at least one carbon-carbon double bond having 2 to 6 carbon atoms. If present at the end of the molecule, examples are CH=CH ₂ , -CH=CH-CH ₃ , -CH ₂ -CH=CH ₂ , -CH=CHCH ₂ -CH ₃ and -CH=CH-CH=CH ₂ . When two constituent molecules are linked by a C _2-6 alkenyl group, then an example of such a C _2-6 alkenyl is -CH=CH-. Each C _2-6 hydrogen atom of the alkenyl moiety may optionally be substituted with a substituent as defined above. Optionally, C _2-6 alkenyl may be interrupted by one or more moieties as defined below.

Accordingly, the term "C _2-10 alkenyl", "C _2-20 alkenyl", or "C _2-50 alkenyl" alone or in combination means a straight or branched hydrocarbon moiety containing at least one carbon-carbon double bond, having 2 to 10, 2 to 20 or 2 to 50 carbon atoms. Each hydrogen atom of a C _2-10 alkenyl, C _2-20 alkenyl, or C _2-50 alkenyl group may optionally be substituted with a substituent as defined above. Optionally, C _2-10 alkenyl, C _2-20 alkenyl, or C _2-50 alkenyl may be interrupted by one or more moieties as defined below.

As used herein, the term "C _2-6 alkynyl" alone or in combination means a straight or branched hydrocarbon moiety containing at least one carbon-carbon triple bond having 2 to 6 carbon atoms. If present at the end of the molecule, examples are -C≡CH, -CH ₂ -C≡CH, CH ₂ -CH ₂ -C≡CH and CH ₂ -C≡C-CH ₃ . When two constituent molecules are linked by an alkynyl group, then -C≡C- is an example. Each C _2-6 hydrogen atom of the alkynyl group may optionally be substituted with a substituent as defined above. Optionally, one or more double bonds may be present. Optionally, C _2-6 alkynyl may be interrupted by one or more moieties as defined below.

Accordingly, as used herein, the terms "C _2-10 alkynyl", "C _2-20 alkynyl" and "C _2-50 alkynyl" alone or in combination mean a straight or branched hydrocarbon moiety containing at least one a carbon-carbon triple bond having 2 to 10, 2 to 20, or 2 to 50 carbon atoms, respectively. Each hydrogen atom of the C _2-10 alkynyl, C _2-20 alkynyl, or C _2-50 alkynyl group may optionally be substituted with a substituent as defined above. Optionally, one or more double bonds may be present. Optionally, C _2-10 alkynyl, C _2-20 alkynyl, or C _2-50 alkynyl may be interrupted by one or more moieties as defined below.

As mentioned above, C _1-4 alkyl, C _1-6 alkyl, C _1-10 alkyl, C _1-20 alkyl, C _1-50 alkyl, C _2-6 alkenyl, C _2-10 alkenyl, C _2-20 alkenyl, C _2-50 alkenyl, C _2-6 alkynyl, C _2-10 alkynyl, C _2-20 alkenyl, or C _2-50 alkynyl may optionally be interrupted by one or more moieties which are preferably selected from the group consisting of

where

dotted lines denote attachment to the remainder of the constituent or reactant; and

-R and R ^a are independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

As used in this application, the term "C _3-10 cycloalkyl" means a cyclic alkyl chain having from 3 to 10 carbon atoms, which may be saturated or unsaturated, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl. Each C _3-10 hydrogen atom of the cycloalkyl carbon may be substituted with a substituent as defined above. The term "C _3-10 cycloalkyl" also includes bridged bicycles such as norbornane or norbornene.

The term "8-30 membered carbopolycyclyl" or "8-30 membered carbopolycycle" means a cyclic moiety of two or more rings with 8 to 30 ring atoms, where two adjacent rings share at least one ring atom, and which may contain up to the maximum number of double bonds (aromatic or non-aromatic ring that is fully saturated, partially unsaturated or unsaturated). Preferably 8-30 membered carbopolycyclyl means a cyclic moiety of two, three, four or five rings, more preferably two, three or four rings.

As used in this application, the term "3-10 membered heterocyclyl" or "3-10 membered heterocycle" means a ring with 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms, which may contain up to the maximum number of double bonds (aromatic or non-aromatic ring that is fully saturated, partially unsaturated or unsaturated), where at least one ring atom to four ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O )-, -S(O) ₂ -), oxygen and nitrogen (including ═N(O)-), and where the ring is linked to the rest of the molecule via a carbon or nitrogen atom. Examples of 3-10 membered heterocycles include, but are not limited to, aziridine, oxirane, tyran, azirine, oxirene, thiirene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazol, isothiazoline, thiadiazole, thiadiazolin, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, imimide, tetra, dihydropyranpyran pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine or homopiperazine. Each hydrogen atom of a 3-10 membered heterocyclyl or 3-10 membered heterocyclic group may be substituted with a substituent as defined below.

As used in this application, the term "8-11 membered heterobicyclyl" or "8-11 membered heterobicycle" means a heterocyclic moiety of two rings with 8 to 11 ring atoms, where at least one ring atom is shared between two rings , and which contains up to the maximum number of double bonds (aromatic or non-aromatic ring that is fully saturated, partially unsaturated or unsaturated), where at least one ring atom to six ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O) ₂ -), oxygen and nitrogen (including=N(O)-), and where the ring is linked to the rest of the molecule via a carbon or nitrogen atom. Examples of 8-11 membered heterobicycles are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazol, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, , benzazepine, purine or pteridine. The term 8-11 membered heterobicycle also includes two-ring spiro structures such as 1,4-dioxa-8-azaspiro[4.5]decane or bridged heterocycles such as 8-aza-bicyclo[3.2.1]octane. Each hydrogen atom of an 8-11 membered heterobicyclyl or an 8-11 membered heterobicycle may be substituted with a substituent as defined below.

Similarly, the term "8-30 membered heteropolycyclyl" or "8-30 membered heteropolycycle" means a heterocyclic moiety of more than two rings with 8 to 30 ring atoms, preferably three, four or five rings, where two adjacent rings share at least one ring atom, and which may contain up to a maximum number of double bonds (aromatic or non-aromatic ring that is fully saturated, partially unsaturated or unsaturated), where at least one ring atom to 10 ring atoms are replaced by a heteroatom selected from the group , consisting of sulfur (including -S(O)-, -S(O) ₂ -), oxygen and nitrogen (including =N(O)-), and where the ring is connected to the rest of the molecule through a carbon or nitrogen atom.

It is understood that the phrase “a pair of R ^x /R ^y joins together with the atom to which they are attached to form a C _3-10 cycloalkyl or 3-10 membered heterocyclyl” in relation to the constituent structure

means that Rx and Ry form the following structure:

,

,

where R is C _3-10 cycloalkyl or 3-10 membered heterocyclyl.

It is also understood that the phrase "a pair of R ^x /R ^y joins together with the atom to which they are attached to form ring A" in relation to the constituent structure

means that R ^x and R ^y form the following structure:

.

.

As used in this application, "halogen" means fluorine, chlorine, bromine or iodine. In general, it is preferred that the halogen is fluorine or chlorine.

In general, the term "comprises" or "comprising" also encompasses "consists of" or "consisting of".

It is clear that human experiments are subject to strict rules. Thus, a more readily available animal model testing system may be needed to determine the dosage of PTH 1-84 required to maintain calcium levels within normal levels in subjects with hypocalcemia, which in humans preferably refers to an albumin-adjusted calcium level above 8.5 mg/day. dl and below 10.5 mg/dl with an optimal level of 9.5 mg/dl, which corresponds to a range of 2.125 to 2.625 nmol/l, with an optimal value of 2.375 nmol/l. One such animal model is thyroparathyroidectomy (TPTX) rats. Rats subjected to thyroparathyroidectomy are unable to produce parathyroid hormone, PTH, the main regulator of calcium homeostasis, and hence will develop hypocalcemia.

However, it is also understood that serum calcium levels can vary between species, such as humans and rats, so values need to be adjusted to account for these species differences in order to achieve comparable results. Therefore, when using such animal models, it is necessary to first determine the normal serum calcium levels for a given sex and age. For example, Watchorn (Biochem J. 1933; 27(6): 1875–1878) provides the normal range of serum calcium (uncorrected) for male rats: 10.29 to 13.16 mg/dL and 9.61 to 14 04 mg/dl for female rats. Preferably normal range of serum calcium is defined as serum calcium concentration above the lower normal range, e.g. as above 9.6 mg/dl (uncorrected) for female rats, even more preferably for female rats between 13 and 22 weeks of age. Preferably, the normal range of serum calcium is defined as a serum calcium concentration below the upper normal range, eg, as low as 14 mg/dl (unadjusted) for female rats, even more preferably for female rats between 13 and 22 weeks of age.

Accordingly, when subjecting female rats to thyroparathyroidectomy to produce TPTX rats, the normal range for treatment target exposure should be above 9.6 mg/dL (no adjustment) and below 14 mg/dL (without adjustment) in such animals aged 13 to 22 weeks.

Preferably, the pharmaceutical composition of the present invention is for use in the treatment of a condition that can be treated with PTH.

The pharmaceutical composition for use according to the present invention is administered no more than once every 24 hours, such as every 24 hours, every 48 hours, every 72 hours, every 96 hours, every 120 hours, every 144 hours, once a week, one once every two weeks. Preferably, the administration of the pharmaceutical composition according to the present invention takes place in multiples of 24 hours, i.e. every N times 24 hours, while N is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 and 14.

In one embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every 24 hours.

In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every 48 hours.

In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every 72 hours.

In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every 96 hours.

In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every 120 hours.

In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every 144 hours.

In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered once every week.

According to the present invention, PTH 1-84 is administered every 24 hours. This means that if the pharmaceutical composition containing the controlled release PTH compound is administered every 24 hours, both the pharmaceutical composition containing the controlled release PTH compound and PTH 1-84 are administered at the same frequency. If the pharmaceutical composition containing the controlled release PTH compound is administered at intervals longer than 24 hours, PTH 1-84 is administered more than once in the interval between two consecutive administrations of the pharmaceutical composition containing the controlled release PTH compound. In such a case, the total amount of PTH1-84 is calculated from all administrations given in the indicated interval between two successive administrations of a pharmaceutical composition containing a controlled release PTH compound of the present invention to provide a basis for determining the molar equivalent dose of the controlled release PTH compound.

The pharmaceutical composition for use according to the present invention is administered by injection. In one embodiment of the present invention, administration is by intramuscular injection. In another embodiment of the present invention, administration is by intravenous injection. In another embodiment of the present invention, administration is via subcutaneous administration.

It is understood that the administration method of the controlled release PTH compound and the administration method of PTH 1-84 are identical, that is, if PTH 1-84 is administered by subcutaneous injection, the controlled release PTH compound is also administered by subcutaneous injection. If PTH 1-84 is administered by intramuscular injection, the controlled release PTH compound is also administered by intramuscular injection.

In one embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered via a syringe. In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered via a pen. In another embodiment of the present invention, the pharmaceutical composition for use according to the present invention is administered using an automatic medical syringe.

In the present invention, a pharmaceutical composition for use according to the present invention is administered no more frequently than every 24 hours with a controlled release PTH compound dose that corresponds to no more than 70% of the molar equivalent dose of PTH 1-84 required to maintain calcium levels within the normal range, preferably above 8.5 mg/dl in humans over a 24 hour period. Preferably, the pharmaceutical composition for use according to the present invention is administered with a dose of the controlled release PTH compound that corresponds to no more than 65%, more preferably with a dose of the controlled release PTH compound that corresponds to no more than 60%, even more preferably with a dose of the controlled release PTH compound release that corresponds to no more than 55%, even more preferably with a dose of the controlled release PTH compound that corresponds to no more than 50%, even more preferably with a dose of the controlled release PTH compound that corresponds to no more than 45%, even more preferably with with a dose of the controlled release PTH compound that corresponds to no more than 40%, even more preferably with a dose of the controlled release PTH compound that corresponds to no more than 35%, and most preferably with a dose of the controlled release PTH compound that rai corresponds to no more than 30% of the PTH 1-84 molar equivalent dose required to maintain calcium levels within the normal range, preferably above 8.5 mg/dl, in humans over a 24 hour period.

Preferably, the PTH compound contains a PTH molecule or PTH moiety having the sequence SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO :112, SEQ ID NO:113, SEQ ID NO:114, or SEQ ID NO:115. More preferably, the PTH molecule or PTH moiety has the sequence SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:110, SEQ ID NO:111, or SEQ ID NO:112.

In one embodiment of the present invention, the PTH molecule or PTH moiety has the sequence of SEQ ID NO:50.

In another embodiment of the present invention, the PTH molecule or PTH moiety has the sequence of SEQ ID NO:52.

In another embodiment of the present invention, the PTH molecule or PTH moiety has the sequence of SEQ ID NO:110.

In another embodiment of the present invention, the PTH molecule or PTH moiety has the sequence of SEQ ID NO:111.

In another embodiment of the present invention, the PTH molecule or PTH moiety has the sequence of SEQ ID NO:112.

Most preferably, the PTH molecule or PTH moiety has the sequence of SEQ ID NO:51.

In one embodiment of the present invention, the PTH controlled release compound is water insoluble.

Preferably, the water insoluble PTH controlled release compound is selected from the group consisting of crystals, nanoparticles, microparticles, nanospheres and microspheres.

In one embodiment of the present invention, the water insoluble controlled release PTH compound is a crystal containing at least one PTH molecule or PTH moiety.

In another embodiment of the present invention, the water insoluble controlled release PTH compound is a nanoparticle containing at least one PTH molecule or PTH moiety.

In another embodiment of the present invention, the water insoluble controlled release PTH compound is a microparticle containing at least one PTH molecule or PTH moiety.

In another embodiment of the present invention, the water insoluble controlled release PTH compound is a nanosphere containing at least one PTH molecule or PTH moiety.

In another embodiment of the present invention, the water-insoluble controlled release PTH compound is a microsphere containing at least one PTH molecule or PTH moiety.

In one embodiment of the present invention, the water-insoluble controlled release PTH compound is a vesicle containing at least one PTH molecule or PTH moiety. Preferably, such a vesicle containing at least one PTH molecule or PTH moiety is a micelle, liposome, or polymersome.

In one embodiment of the present invention, the water insoluble controlled release PTH compound is a micelle containing at least one PTH molecule or PTH moiety.

In another embodiment of the present invention, the water insoluble controlled release PTH compound is a liposome containing at least one PTH molecule or PTH moiety. Preferably, such liposome is selected from the group consisting of aquasomes; non-ionic surface active vesicles such as niosomes and proniosomes; cationic liposomes such as LeciPlex; transfersomes; ethosomes; ufasomes; sphingosomes; and pharmacosomes.

In another embodiment of the present invention, the water insoluble controlled release PTH compound is a polymersome containing at least one PTH molecule or PTH moiety.

In another embodiment of the present invention, the water-insoluble controlled release PTH compound comprises at least one PTH molecule non-covalently incorporated into the water-insoluble polymer. Preferably, such a water-insoluble polymer comprises a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphoylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy)polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly (dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl- oxazolines), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-glycolic acid copolymers), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly (organophosphazenes), poly(orthoesters), poly(oxazolines), poly(propyl glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinylmethyl ethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethylcelluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans , dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other polymers based on carbohydrates, xylans and their copolymers.

In a preferred embodiment of the present invention, the water-insoluble controlled release PTH compound comprises at least one PTH molecule non-covalently incorporated into a poly(lactic-glycolic acid copolymer) (PLGA).

In another embodiment of the present invention, the water insoluble controlled release PTH compound contains at least one PTH moiety covalently and reversibly conjugated to the water insoluble polymer. Preferably, such a water-insoluble polymer comprises a polymer comprising a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphoylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides) , poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl -oxazolines), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazolines), poly(iminocarbonates), poly(lactic acids), poly(copolymers of lactic and glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(orthoesters), poly(oxazolino c), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinylmethyl ethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethylcelluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other polymers based on carbohydrates, xylans and their copolymers.

In one embodiment of the present invention, such a controlled release water-insoluble PTH compound is a compound containing a D-L conjugate, where

-D is the PTH component; and

-L contains a reversible prodrug linker moiety -L ¹ -, where the -L ¹ - moiety is connected to the PTH moiety of -D via a PTH moiety;

where -L ¹ - is substituted with -L ² -Z' and is optionally further substituted; where

-L ² - represents a simple chemical bond or spacer component; and

-Z' is the water-insoluble component of the carrier.

It is understood that a plurality of -L ² -L ¹ -D moieties are coupled to a water-insoluble carrier -Z', and that such controlled release PTH compounds are PTH prodrugs more specifically associated with a PTH prodrug carrier.

Preferred options for -D, -L ¹ -, -L ² - and -Z' are described below.

In a preferred embodiment of the present invention, the controlled release PTH compound is water soluble.

In a preferred embodiment of the present invention, such a controlled release water-soluble PTH compound is a compound of formula (Ia) or (Ib) or a pharmaceutically acceptable salt thereof.

(Ia)

(Ia)

(Ib),

(Ib),

where

-D is the PTH component;

-L ¹ - is a reversible prodrug linker moiety reversibly or covalently linked to the PTH moiety of -D via a PTH moiety;

-L ² - represents a simple chemical bond or spacer component;

-Z is the water-soluble component of the carrier;

x is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16; and

y is an integer selected from the group consisting of 1, 2, 3, 4 and 5.

It is understood that the compounds of formula (Ia) and (Ib) are PTH prodrugs, more particularly water-soluble PTH prodrugs.

Preferably, -D has the sequence SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO: 113, SEQ ID NO:114 or SEQ ID NO:115. More preferably, -D has the sequence SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:110, SEQ ID NO:111, or SEQ ID NO:112.

In one embodiment of the present invention, -D has the sequence of SEQ ID NO:50.

In another embodiment of the present invention, -D has the sequence of SEQ ID NO:52.

In another embodiment of the present invention, -D has the sequence of SEQ ID NO:110.

In another embodiment of the present invention, -D has the sequence of SEQ ID NO:111.

In another embodiment of the present invention, -D has the sequence of SEQ ID NO:112.

Most preferably, -D has the sequence of SEQ ID NO:51.

The -L ¹ - moiety is either conjugated to the side chain functional group of the amino acid residue -D, to the N-terminal amine functional group or to the C-terminal carboxyl functional group -D, or to the nitrogen atom of the main polypeptide chain -D. Attachment to either the N-terminus or the C-terminus can be either directly via the appropriate amine or carboxyl functional group, respectively, or indirectly, when the spacer moiety is first conjugated to the amine or carboxyl functional group to which the -L ¹ spacer moiety is conjugated -.

Preferably, the PTH amino acid residue to which -L ¹ is conjugated contains a functional group selected from the group consisting of carboxylic acid, primary and secondary amine, maleimide, thiol, sulfonic acid, carbonate, carbamate, hydroxyl, aldehyde, ketone, hydrazine , isocyanate, isothiocyanate, phosphoric acid, phosphonic acid, haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, sulfate, disulfide, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, guanidine, and aziridine. Even more preferably, the PTH amino acid residue to which -L ¹ - is conjugated contains a functional group selected from the group consisting of hydroxyl, primary and secondary amine, and guanidine. Even more preferably, the PTH amino acid residue to which -L ¹ - is conjugated contains a primary or secondary amine functional group. Most preferably, the PTH amino acid residue to which -L ¹ - is conjugated contains a primary amine functional group.

When the -L ¹ - moiety is conjugated to a PTH amino acid residue side chain functional group, said amino acid residue is selected from the group consisting of proteogenic amino acid residues and non-proteogenic amino acid residues.

In one embodiment of the present invention, -L ¹ - is conjugated to a side chain functional group of a non-proteogenic PTH amino acid residue. It is understood that such a non-proteogenic amino acid is not found in the sequence of native PTH or fragments thereof, and that it can only be present in variants and derivatives of PTH.

In another embodiment of the present invention, -L ¹ - is conjugated to a side chain functional group of a proteogenic acid residue in PTH. Preferably, said amino acid is selected from the group consisting of histidine, lysine, tryptophan, serine, threonine, tyrosine, aspartic acid, glutamic acid and arginine. Even more preferably, said amino acid is selected from the group consisting of lysine, aspartic acid, arginine and serine. Even more preferably, said amino acid is selected from the group consisting of lysine, arginine and serine.

In one embodiment of the present invention, -L ¹ - is conjugated to a PTH histidine side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH lysine side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH tryptophan side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH serine side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH threonine side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH tyrosine side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH aspartic acid side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH glutamic acid side chain functional group.

In another embodiment of the present invention, -L ¹ - is conjugated to a PTH arginine side chain functional group.

It is understood that not every PTH moiety may contain all of these amino acid residues.

In a preferred embodiment of the present invention, -L ¹ - is conjugated to the N-terminal amine functionality of PTH, either directly via the corresponding amine functionality, or indirectly when the spacer moiety is first conjugated to the amine functionality to which the spacer moiety -L ¹ is conjugated -. Even more preferably, -L ¹ - is directly conjugated to the N-terminal amine functional group of PTH.

In an equally preferred embodiment of the present invention, --L ¹ - is conjugated to the C-terminal PTH functionality, either directly via the corresponding carboxyl functionality, or indirectly when the spacer moiety is first conjugated to the carboxyl functionality to which the -L ¹ spacer moiety is conjugated -.

Most preferably, L ¹ is directly conjugated to the N-terminal amine functionality of PTH.

Component -L ¹ - can be connected to -D through any type of connection, provided that it is reversible. Preferably, -L ¹ - is connected to -D via a bond selected from the group consisting of amide, ester, carbamate, acetal, aminal, imine, oxime, hydrazone, disulfide and acylguanidine. Even more preferably -L ¹ - is connected to -D through a bond selected from the group consisting of amide, ester, carbamate and acylguanidine. It is understood that some of these bonds are not themselves reversible, but in the present invention, the neighboring groups contained in L ¹ make these bonds reversible.

In one embodiment of the present invention, -L ¹ - is connected to -D via an ether bond.

In another embodiment of the present invention, -L ¹ - is connected to -D via a carbamate bond.

In another embodiment of the present invention, -L ¹ - is connected to -D via an acylguanidine bond.

In a preferred embodiment of the present invention, -L ¹ - is connected to -D via an amide bond.

The -L ¹ - moiety is a reversible prodrug linker from which the drug, ie. PTH is released in its free form, i.e. is a traceless prodrug linker. Suitable prodrug linkers are known in the art, such as reversible prodrug linker moieties, are disclosed in WO 2005/099768 A2, WO 2006/136586 A2, WO 2011/089216 A1 and WO 2013/024053 A1, which are incorporated herein by reference.

In another embodiment of the present invention -L ¹ - is a reversible prodrug linker as described in WO 2011/012722 A1, WO 2011/089214 A1, WO 2011/089215 A1, WO 2013/024052 A1 and WO 2013/160340 A1, which incorporated into this application by reference.

A particularly preferred component -L ¹ - is disclosed in WO 2009/095479 A2. Accordingly, in a preferred embodiment of the present invention, the -L ¹ - component has the formula (II):

,

,

where the dotted line denotes accession to the nitrogen atom, hydroxyl or thiol of the component -D, which is the PTH component;

-X- is selected from the group consisting of -C(R ⁴ R ^4a )-; -N(R ⁴ )-; -O-; -C(R ⁴ R ^4a )-C(R ⁵ R ^5a )-; -C(R ⁵ R ^5a )-C(R ⁴ R ^4a )-; -C(R ⁴ R ^4a )-N(R ⁶ )-; -N(R ⁶ )-C(R ⁴ R ^4a )-; -C(R ⁴ R ^4a )-O-; -OC(R ⁴ R ^4a )-; and -C(R ⁷ R ^7a )-;

X ¹ is selected from the group consisting of C; and S(O);

-X ² - selected from the group consisting of -C(R ⁸ R ^8a )-; and -C(R ⁸ R ^8a )-C(R ⁹ R ^9a )-;

=X ³ is selected from the group consisting of=O;=S; u=N-CN;

-R ¹ , -R ^1a , -R ² , -R ^2a , -R ⁴ , -R ^4a , -R ⁵ , -R ^5a , -R ⁶ , -R ⁸ , -R ^8a , -R ⁹ , and - R ^9a are independently selected from the group consisting of -H; and C _1-6 alkyl;

-R ³ and -R ^3a are independently selected from the group consisting of -H; and C _1-6 alkyl, with the proviso that if one of -R ³ , -R ^3a or both is other than hydrogen, they are connected to the N to which they are attached via an SP ³ hybridized carbon atom;

-R ⁷ is selected from the group consisting of -N(R ¹⁰ R ^10a ); and -NR ¹⁰ -(C=O) -R ¹¹ ;

-R ^7a , -R ¹⁰ , -R ^10a , and -R ¹¹ are independently selected from the group consisting of -H; and C _1-6 alkyl;

optionally, one or more of the pairs -R ^1a /-R ^4a , -R ^1a /-R ^5a , -R ^1a /-R ^7a , -R ^4a /-R ^5a , and -R ^8a /-R ^9a form a chemical bond ;

optionally, one or more of the pairs -R ¹ /-R ^1a , -R ² /-R ^2a , -R ⁴ /-R ^4a , -R ⁵ /-R ^5a , -R ⁸ /-R ^8a ,

and -R ⁹ /-R ^9a are connected together with the atom to which they are attached, with the formation of C _3-10 cycloalkyl; or 3-10 membered heterocyclyl;

optionally, one or more of the pairs -R ¹ /-R ⁴ , -R ¹ /-R ⁵ , -R ¹ /-R ⁶ , -R ¹ /-R ^7a , -R ⁴ /-R ⁵ , -R ⁴ /-R ⁶ , -R ⁸ /-R ⁹ , and -R ² /-R ³ join together with the atoms to which they are attached to form ring A;

optionally, R ³ /R ^3a join together with the nitrogen atom to which they are attached to form a 3-10 membered heterocycle;

A is selected from the group consisting of phenyl; naphthyl; indenyl; indanyl; tetralinyl; C _3-10 cycloalkyl; 3-10 membered heterocyclyl; and 8-11 membered heterobicyclyl; and

where -L ¹ - is substituted with -L ² -Z or -L ² -Z', and where -L ¹ - is optionally further substituted, provided that the asterisked hydrogen in formula (II) is not substituted with -L ² -Z or -L ² -Z' or a substituent;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Preferably -L ¹ - of formula (II) has as a substituent one component -L ² -Z or -L ² -Z'.

In one embodiment of the present invention, -L ¹ - of formula (II) is not further substituted.

It is clear that if -R ³ /-R ^3a of formula (II) are connected together with the nitrogen atom to which they are attached to form a 3-10 membered heterocycle, only such 3-10 membered heterocycles can be formed, in in which the atoms directly attached to the nitrogen atom are SP ³ hybridized carbon atoms. In other words, such a 3-10 membered heterocycle formed by -R ³ /-R ^3a together with the nitrogen atom to which they are attached has the following structure:

where

the dotted line denotes attachment to the remainder -L ¹ -;

the ring contains from 3 to 10 atoms, including at least one nitrogen atom; and

R ^# and R ^## represent SP ³ -hybridized carbon atom.

It is also understood that a 3 to 10 membered heterocycle may be further substituted.

Exemplary embodiments of suitable 3 to 10 membered heterocycles formed by -R ³ /-R ^3a of formula (II) together with the nitrogen atom to which they are attached are as follows:

where

the dotted line denotes attachment to the remainder of the molecule; and

-R is selected from the group consisting of -H and C _1-6 alkyl.

-L ¹ - of formula (II) may optionally be further substituted. In general, any substituent may be used, as long as the principle of cleavage is not affected, i.e. the hydrogen marked with an asterisk in formula (II) is not replaced, and the nitrogen of the constituent

formula (II) remains part of the primary, secondary or tertiary amine, i.e. -R ³ and R ^3a independently of each other represent -H or connected to -N< through SP ³ -hybridized carbon atom.

In one embodiment of the present invention, -R ¹ or R ^1a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ² or R ^2a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ³ or R ^3a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ⁴ of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ⁵ or R ^5a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ⁶ of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ⁷ or R ^7a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ⁸ or R ^8a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ⁹ or R ^9a of formula (II) is substituted with -L ² -Z or L ² -Z'. In another embodiment of the present invention, -R ¹⁰ is substituted with -L ² -Z or -L ² -Z'. In another embodiment of the present invention, -R ¹¹ is substituted with -L ² -Z or -L ² -Z'.

Preferably, -X- of formula (II) is selected from the group consisting of -C(R ⁴ R ^4a )-, -N(R ⁴ )- and -C(R ⁷ R ^7a )-.

In one embodiment of the present invention, -X- of formula (II) is -C(R ⁴ R ^4a )-.

In one preferred embodiment of the present invention, the -X- of formula (II) is -C(R ⁷ R ^7a )-.

Preferably, -R ⁷ of formula (II) is -NR ¹⁰ -(C=O)-R ¹¹ .

Preferably, -R ^7a of formula (II) is selected from H, methyl and ethyl. Most preferably -R ^7a of formula (II) is -H.

Preferably, -R ¹⁰ is selected from H, methyl and ethyl. Most preferably -R ¹⁰ is methyl.

Preferably, -R ¹¹ is selected from H, methyl and ethyl. Most preferably -R ¹¹ is -H.

Preferably, -R ¹¹ is substituted with -L ² -Z or -L ² -Z'.

In another preferred embodiment of the present invention, -X- of formula (II) is -N(R ⁴ )-.

Preferably, -R ⁴ is selected from the group consisting of H, methyl and ethyl. Preferably, -R ⁴ is -H.

Preferably X ^{1 of} formula (II) is C.

Preferably, ═X ^{3 of} formula (II) is ═O.

Preferably, -X ² - of formula (II) is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and -R ^8a of formula (II) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ⁸ and -R ^8a of formula (II) is -H. Even more preferably, both -R ⁸ and -R ^8a of formula (II) are -H.

Preferably, -R ¹ and -R ^1a of formula (II) are independently selected from the group consisting of H, methyl and ethyl.

In one preferred embodiment of the present invention, at least one of -R ¹ and -R ^1a of formula (II) is -H, more preferably both -R ¹ and -R ^1a of formula (II) are -H.

In another preferred embodiment of the present invention, at least one of -R ¹ and -R ^1a of formula (II) is methyl, more preferably both -R ¹ and R ^1a of formula (II) are -methyl.

Preferably, -R ² and R ^2a of formula (II) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ² and R ^2a of formula (II) is -H. Even more preferably, both -R ² and R ^2a of formula (II) are -H.

Preferably, -R ³ and R ^3a of formula (II) are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl.

In one preferred embodiment of the present invention, at least one of -R ³ and -R ^3a of formula (II) is methyl, more preferably -R ^{3 of} formula (II) is methyl and R ^3a of formula (II) is -H .

In another preferred embodiment of the present invention, -R ³ and -R ^3a of formula (II) are both -H.

Preferably, -D is connected to -L ¹ - through the nitrogen atom through the formation of an amide bond.

In one preferred embodiment of the present invention, the component -L ¹ - has the formula (IIa-i):

(IIa-i),

(IIa-i),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

-R ¹ , -R ^1a , -R ² , -R ^2a , -R ³ , -R ^3a , -R ⁷ , -R ^7a and -X ² - have the meanings as defined in formula (II); and

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIa-i) is not substituted with -L ² -Z or -L ² -Z' or a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIa-i) or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably -L ¹ - of formula (IIa-i) is substituted with one component -L ² -Z or -L ² -Z'.

Preferably, the -L ¹ - component of formula (IIa-i) is not further substituted.

Preferably, -R ¹ and R ^1a of formula (IIa-i) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ¹ and -R ^1a of formula (IIa-i) is -H. Even more preferably, both -R ¹ and -R ^1a of formula (IIa-i) are -H.

Preferably, -R ⁷ of formula (IIa-i) is -NR ¹⁰ -(C=O)-R ¹¹ .

Preferably, -R ^7a of formula (II-i) is selected from H, methyl and ethyl. Most preferably -R ^7a of formula (II-i) is -H.

Preferably, -R ¹⁰ of formula (IIa-i) is selected from H, methyl and ethyl. Most preferably -R ¹⁰ of formula (IIa-i) is methyl.

Preferably, -R ¹¹ of formula (IIa-i) is selected from H, methyl and ethyl. Most preferably -R ¹¹ of formula (IIa-i) is -H.

Preferably, -R ¹¹ of formula (IIa-i) is substituted with -L ² -Z or L ² -Z'.

Preferably, -X ² - of formula (IIa-i) is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and -R ^8a of formula (IIa-i) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ⁸ and -R ^8a of formula (IIa-i) is -H. Even more preferably, both -R ⁸ and -R ^8a of formula (IIa-i) are -H.

Preferably, -R ² and -R ^2a of formula (IIa-i) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ² and -R ^2a of formula (IIa-i) is -H. Even more preferably, both -R ² and -R ^2a of formula (IIa-i) are -H.

Preferably, -R ³ and -R ^3a of formula (IIa-i) are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Even more preferably, at least one of -R ³ and -R ^3a of formula (IIa-i) is methyl.

Preferably, -R ^{3 of} formula (IIa-i) is -H and -R ^3a of formula (IIa-i) is methyl.

More preferably, the -L ¹ - component has the formula (IIa-ii):

(IIa-ii),

(IIa-ii),

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

-R ² , -R ^2a , -R ¹⁰ , -R ¹¹ and -X ² - have the meanings as defined in formula (II); and

where -L ¹ - is substituted with -L ² -Z or L ² -Z' and where -L ¹ - is optionally further substituted, provided that the asterisked hydrogen in formula (IIa-ii) is not substituted with -L ² -Z or -L ² -Z' or a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIa-ii) or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably -L ¹ - of formula (IIa-ii) is substituted with one component -L ² -Z or -L ² -Z'.

Preferably, the -L ¹ - component of formula (IIa-ii) is not further substituted.

Preferably, -X ² - of formula (IIa-ii) is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and -R ^8a of formula (IIa-ii) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ⁸ and R ^8a of formula (IIa-ii) is -H. Even more preferably, both -R ⁸ and R ^8a of formula (IIa-ii) are -H.

Preferably, -R ³ and R ^3a of formula (IIa-ii) are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Even more preferably, at least one of -R ³ and R ^3a of formula (IIa-ii) is methyl.

Preferably, -R ^{3 of} formula (IIa-ii) is -H and R ^3a of formula (IIa-ii) is methyl.

Preferably, -R ¹⁰ of formula (IIa-ii) is selected from H, methyl and ethyl. Most preferably -R ¹⁰ of formula (IIa-ii) is methyl.

Preferably, -R ¹¹ of formula (IIa-ii) is selected from H, methyl and ethyl. Most preferably -R ¹¹ of formula (IIa-ii) is -H.

Preferably, -R ¹¹ of formula (IIa-ii) is substituted with -L ² -Z or L ² -Z'.

In an even more preferred embodiment of the present invention, the component -L ¹ - has the formula (IIa-ii'):

(IIa-ii’),

(IIa-ii'),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

dashed line marked with an asterisk means attachment to -L ² -;

-R ³ , -R ^3a , -R ¹⁰ and X ² - are as defined in formula (II); and

where -L ¹ - optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIa-ii') is not substituted by a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIa-ii') or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably, the -L ¹ - component of formula (IIa-ii') is not further substituted.

Preferably, -X ² - of formula (IIa-ii') is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and R ^8a of formula (IIa-ii') are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ⁸ and R ^8a of formula (IIa-ii') is -H. Even more preferably, both -R ⁸ and R ^8a of formula (IIa-ii') are -H.

Preferably, -R ³ and R ^3a of formula (IIa-ii') are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Even more preferably, at least one of -R ³ and R ^3a of formula (IIa-ii') is methyl.

Preferably, -R ^{3 of} formula (IIa-ii') is -H and R ^3a of formula (IIa-ii') is methyl.

Preferably, -R ¹⁰ of formula (IIa-ii') is selected from H, methyl and ethyl. Most preferably -R ¹⁰ of formula (IIa-ii') is methyl.

Even more preferably, the -L ¹ - component has the formula (IIa-iii):

(IIa-iii),

(IIa-iii),

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond; and

where -L ¹ - is substituted with -L ² -Z or -L ² -Z', and where -L ¹ - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIa-iii) is not substituted with -L ² - Z or -L ² -Z' or a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIa-iii) or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably -L ¹ - of formula (IIa-iii) is substituted with one component -L ² -Z or -L ² -Z'.

Preferably, the -L ¹ - component of formula (IIa-iii) is not further substituted.

Most preferably, the -L ¹ - component has the formula (IIa-iii'):

(IIa-iii’),

(IIa-iii'),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

dashed line marked with an asterisk means attachment to -L ² -;

-R ² , -R ^2a , -R ³ , -R ^3a and X ² - are as defined in formula (II); and

where -L ¹ - optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIa-iii') is not substituted by a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIa-iii') or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably, the -L ¹ - component of formula (IIa-iii') is not further substituted.

In another preferred embodiment of the present invention, the component -L ¹ - has the formula (IIb-i)

(IIb-i),

(IIb-i),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

-R ¹ , -R ^1a , -R ² , -R ^2a , -R ³ , -R ^3a , -R ⁴ and X ² - are as defined in formula (II); and

where -L ¹ - is substituted with -L ² -Z or -L ² -Z', and where -L ¹ - is optionally further substituted, provided that the asterisked hydrogen in formula (IIb-i) is not substituted with -L ² - Z or -L ² -Z' or a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIb-i) or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably -L ¹ - of formula (IIb-i) is substituted with one component -L ² -Z or -L ² -Z'.

Preferably, the -L ¹ - component of formula (IIb-i) is not further substituted.

Preferably, -R ¹ and R ^1a of formula (IIb-i) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ¹ and R ^1a of formula (IIb-i) is methyl. Even more preferably, both -R ¹ and R ^1a of formula (IIb-i) are -methyl.

Preferably, -R ⁴ of formula (IIb-i) is selected from the group consisting of H, methyl and ethyl. More preferably, -R ⁴ of formula (IIb-i) is -H.

Preferably, -X ² - of formula (IIb-i) is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and R ^8a of formula (IIb-i) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ⁸ and R ^8a of formula (IIb-i) is -H. Even more preferably, both -R ⁸ and R ^8a of formula (IIb-i) are -H.

Preferably, -R ² and R ^2a of formula (IIb-i) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ² and R ^2a of formula (IIb-i) is -H. Even more preferably, both -R ² and R ^2a of formula (IIb-i) are -H.

Preferably, -R ³ and R ^3a of formula (IIb-i) are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Even more preferably, at least one of -R ³ and R ^3a of formula (IIb-i) is -H. Even more preferably, both -R ³ and R ^3a of formula (IIb-i) are -H.

More preferably, the -L ¹ - component has the formula (IIb-ii):

(IIb-ii),

(IIb-ii),

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

-R ² , -R ^2a , -R ³ , -R ^3a and X ² - are as defined in formula (II); and

where -L ¹ - is substituted with -L ² -Z or -L ² -Z', and where -L ¹ - is optionally further substituted, provided that the asterisked hydrogen in formula (IIb-ii) is not substituted with -L ² - Z or -L ² -Z' or a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a formula (IIb-ii) or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably -L ¹ - of formula (IIb-ii) is substituted with one component -L ² -Z or -L ² -Z'.

Preferably, the -L ¹ - component of formula (IIb-ii) is not further substituted.

Preferably, -X ² - of formula (IIb-ii) is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and R ^8a of formula (IIb-ii) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ⁸ and R ^8a of formula (IIb-ii) is -H. Even more preferably, both -R ⁸ and R ^8a of formula (IIb-ii) are -H.

Preferably, -R ² and R ^2a of formula (IIb-ii) are independently selected from the group consisting of H, methyl and ethyl. More preferably, at least one of -R ² and R ^2a of formula (IIb-ii) is -H. Even more preferably, both -R ² and R ^2a of formula (IIb-ii) are -H.

Preferably, -R ³ and R ^3a of formula (IIb-ii) are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Even more preferably, at least one of -R ³ and R ^3a of formula (IIb-ii) is -H. Even more preferably, both -R ³ and R ^3a of formula (IIb-ii) are -H.

Even more preferably, the -L ¹ - component has the formula (IIb-ii'):

(IIb-ii’),

(IIb-ii'),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

-R ² , -R ^2a , -R ³ , -R ^3a and X ² - are as defined in formula (II); and

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIb-ii') is not substituted with -L ² - Z or -L ² -Z' or a substituent.

It will be understood that if -R ^3a of formula (IIb-ii') is other than -H, it is connected to N to which it is attached via an sp ³ hybridized carbon atom.

Preferably, the -L ¹ - moiety of formula (IIb-ii') is not further substituted.

Preferably, -X ² - of formula (IIb-ii') is -C(R ⁸ R ^8a )-.

Preferably -R ⁸ and -R ^8a of formula (IIb-ii') are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R ⁸ and -R ^8a of formula (IIb-ii') is -H. Even more preferably, both -R ⁸ and -R ^8a of formula (IIb-ii') are -H.

Preferably, -R ² and -R ^2a of formula (IIb-ii') are independently selected from the group consisting of -H, methyl and ethyl. More preferably, at least one of -R ² and -R ^2a of formula (IIb-ii') is -H. Even more preferably, both -R ² and -R ^2a of formula (IIb-ii') are H.

Preferably, -R ^3a of formula (IIb-ii') is selected from the group consisting of -H, methyl, ethyl, propyl and butyl. In one embodiment of the present invention, -R ^3a of formula (IIb-ii') is -H.

Even more preferably, the -L ¹ - component has the formula (IIb-iii):

(IIb-iii),

(IIb-iii),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond; and

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted, provided that the hydrogen marked with an asterisk in formula (IIb-iii) is not substituted with -L ² -Z or L ² -Z' or a substituent.

It is clear that in the case where one of the -R ³ , -R ^3a of the formula (IIb-iii) or both other than -H, they are connected to N, to which they are attached through sp ³ -hybridized carbon atom.

Preferably -L ¹ - of formula (IIb-iii) is substituted with one component -L ² -Z or L ² -Z'.

Preferably, the -L ¹ - component of formula (IIb-iii) is not further substituted.

Most preferably, the -L ¹ - moiety has the formula (IIb-iii'):

(IIb-iii’),

(IIb-iii'),

where

where the dotted line denotes the addition to the nitrogen atom of the D component, which is the PTH component, by forming an amide bond;

the dotted line marked with an asterisk indicates attachment to -L ² -; and

where -L ¹ - is optionally further substituted, provided that the hydrogen atom marked with an asterisk in formula (IIb-iii') is not substituted with -L ² -Z or -L ² -Z' or a substituent.

It is understood that the nitrogen adjacent to the dotted line marked with an asterisk in the formula (IIb-iii') is attached to -L ² - through sp ³ -hybridized carbon atom.

Preferably, the -L ¹ - component of formula (IIb-iii') is not further substituted.

Another preferred moiety, L ¹ , is disclosed in WO2016/020373A1. Accordingly, in another preferred embodiment of the present invention, the -L ¹ - component has the formula (III):

(III),

(III)

where

the dotted line shows attachment to the primary or secondary amine or hydroxyl of the D moiety via amide or ester bond formation, respectively;

-R ¹ , -R ^1a , -R ² , -R ^2a , -R ³ and R ^3a are independently selected from the group consisting of -H, -C(R ⁸ R ^8a R ^8b ), -C(= O)R ⁸ , -C≡N, -C(=NR ⁸ )R ^8a , -CR ⁸ (=CR ^8a R ^8b ), -C≡CR ⁸ and T;

-R ⁴ , -R ⁵ and R ^5a are independently selected from the group consisting of -H, -C(R ⁹ R ^9a R ^9b ) and T;

a1 and a2 are independently 0 or 1;

each -R ⁶ , -R ^6a , -R ⁷ , -R ^7a , -R ⁸ , -R ^8a , -R ^8b , -R ⁹ , -R ^9a , -R ^9b are independently selected from the group consisting of -H, halogen, -CN, -COOR ¹⁰ , -OR ¹⁰ , -C(O)R ¹⁰ , -C(O)N(R ¹⁰ R ^10a ), -S(O) ₂ N(R ¹⁰ R ^10a ) , -S(O)N(R ¹⁰ R ^10a ), -S(O) ₂ R ¹⁰ , -S(O)R ¹⁰ , -N(R ¹⁰ )S(O) ₂ N(R ^10a R ^10b ), -SR ¹⁰ , -N(R ¹⁰ R ^10a ), -NO ₂ , -OC(O)R ¹⁰ , -N(R ¹⁰ )C(O)R ^10a , -N(R ¹⁰ )S(O) ₂ R ^10a , -N(R ¹⁰ )S(O)R ^10a , -N(R ¹⁰ )C(O)OR ^10a , -N(R ¹⁰ )C(O)N(R ^10a R ^10b ), -OC(O )N(R ¹⁰ R ^10a ), -T, C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl; where -T, C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl are optionally substituted with one or more substituents -R ¹¹ that are the same or different, and where C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ¹² )-, -S(O) ₂ N(R ¹² )-, -S(O)N(R ¹² )-, -S(O) ₂ -, -S(O)-, -N(R ¹² ) S(O) ₂ N(R ^12a )-, -S-, -N(R ¹² )-, -OC(OR ¹² )(R ^12a )-, -N(R ¹² )C(O)N(R ^12a )- and -OC(O)N(R ¹² )-;

each -R ¹⁰ , -R ^10a , -R ^10b is independently selected from the group consisting of -H, -T, C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl; where -T, C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl are optionally substituted with one or more substituents -R ¹¹ that are the same or different, and where C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ¹² )-, -S(O) ₂ N(R ¹² )-, -S(O)N(R ¹² )-, -S(O) ₂ -, -S(O)-, -N(R ¹² ) S(O) ₂ N(R ^12a )-, -S-, -N(R ¹² )-, -OC(OR ¹² )(R ^12a )-, -N(R ¹² )C(O)N(R ^12a )- and -OC(O)N(R ¹² )-;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, and 8-11 membered heterobicyclyl; where each T is independently optionally substituted with one or more substituents -R ¹¹ that are the same or different;

each -R ¹¹ is independently selected from halogen, -CN, oxo (=O), -COOR ¹³ , -OR ¹³ , -C(O)R ¹³ , -C(O)N(R ¹³ R ^13a ), -S(O) ₂ N(R ¹³ R ^13a ), -S(O)N(R ¹³ R ^13a ), -S(O) ₂ R ¹³ , -S(O)R ¹³ , -N(R ¹³ ) S(O) ₂ N(R ^13a R ^13b ), -SR ¹³ , -N(R ¹³ R ^13a ), -NO ₂ , -OC(O)R ¹³ , -N(R ¹³ )C(O)R ^13a , -N(R ¹³ )S(O) ₂ R ^13a , -N(R ¹³ )S(O)R ^13a , -N(R ¹³ )C(O)OR ^13a , -N(R ¹³ )C(O )N(R ^13a R ^13b ), -OC(O)N(R ¹³ R ^13a ) and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogen substituents that are the same or different;

each -R ¹² , -R ^12a , -R ¹³ , -R ^13a , -R ^13b is independently selected from the group consisting of -H and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogen substituents that are the same or different;

optionally, one or more of the pairs -R ¹ /-R ^1a , -R ² /-R ^2a , -R ³ /-R ^3a , -R ⁶ /-R ^6a , -R ⁷ /-R ^7a are connected together with the atom to which they are attached to form C _3-10 cycloalkyl or 3-10 membered heterocyclyl;

optionally, one or more of the pairs -R ¹ /-R ² , -R ¹ /-R ³ , -R ¹ /-R ⁴ , -R ¹ /-R ⁵ , -R ¹ /-R ⁶ , -R ¹ /-R ⁷ , -R ² /-R ³ , -R ² /-R ⁴ , -R ² /-R ⁵ , -R ² /-R ⁶ , -R ² /-R ⁷ , -R ³ /- R ⁴ , -R ³ /-R ⁵ , -R ³ /-R ⁶ , -R ³ /-R ⁷ , -R ⁴ /-R ⁵ , -R ⁴ /-R ⁶ , -R ⁴ /-R ⁷ , -R ⁵ /-R ⁶ , -R ⁵ /-R ⁷ , -R ⁶ /-R ⁷ join together with the atoms to which they are attached to form ring A;

A is selected from the group consisting of phenyl; naphthyl; indenyl; indanyl; tetralinyl; C _3-10 cycloalkyl; 3-10 membered heterocyclyl; and 8-11 membered heterobicyclyl;

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Optional other substituents of the -L ¹ - component of formula (III) are preferably as described above.

Preferably -L ¹ - of formula (III) is substituted with one component -L ² -Z or L ² -Z'.

In one embodiment of the present invention, -L ¹ - of formula (III) is not further substituted.

Additional preferred embodiments for -L ¹ - are disclosed in EP1536334B1, WO2009/009712A1, WO2008/034122A1, WO2009/143412A2, WO2011/082368A2 and US8618124B2, which are incorporated herein by reference in their entirety.

Additional preferred embodiments for -L ¹ - are disclosed in US8946405B2 and US8754190B2, which are incorporated herein by reference in their entirety. Accordingly, the preferred component -L ¹ - has the formula (IV):

(IV),

(IV)

where

the dotted line shows attachment to -D, which is the PTH moiety, and where the attachment is through the functional group of the -D moiety selected from the group consisting of -OH, -SH and NH ₂ ;

m is 0 or 1;

at least one or both of -R ¹ and R ² is selected/independently selected from the group consisting of -CN, -NO ₂ , optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, -C (O)R ³ , -S(O)R ³ , -S(O) ₂ R ³ and -SR ⁴ ,

one and only one of -R ¹ and R ² is selected from the group consisting of -H, optionally substituted alkyl, optionally substituted arylalkyl, and optionally substituted heteroarylalkyl;

-R ³ is selected from the group consisting of -H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -OR ⁹ and N(R ⁹ ) ₂ ;

-R ⁴ is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

each -R ⁵ is independently selected from the group consisting of -H, optionally substituted alkyl, optionally substituted alkenylalkyl, optionally substituted alkynylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

-R ⁹ is selected from the group consisting of -H and optionally substituted alkyl;

-Y- is absent and -X- is -O- or S-; or

-Y- is -N(Q)CH ₂ - and X- is -O-;

Q is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

optionally, -R ¹ and R ² can be combined to form a 3-8 membered ring; and

optionally, both -R ⁹ together with the nitrogen atom to which they are attached, form a heterocyclic ring;

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Only in the context of formula (IV) the terms used have the following meanings:

The term "alkyl", as used in this application, includes linear, branched or cyclic saturated hydrocarbon groups having from 1 to 8 carbon atoms, or in some embodiments of the present invention, from 1 to 6 or from 1 to 4 carbon atoms.

The term "alkoxy" includes alkyl groups associated with oxygen, including methoxy, ethoxy, isopropoxy, cyclopropoxy, cyclobutoxy and the like.

The term "alkenyl" includes non-aromatic unsaturated hydrocarbons with carbon-carbon double bonds.

The term "alkynyl" includes non-aromatic unsaturated hydrocarbons with carbon-carbon triple bonds.

The term "aryl" includes aromatic hydrocarbon groups having 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, including groups such as phenyl, naphthyl and anthracenyl. The term "heteroaryl" includes aromatic rings containing from 3 to 15 carbon atoms containing at least one N, O or S atom, preferably from 3 to 7 carbon atoms containing at least one N, O or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isooxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl and the like.

In some cases, alkenyl, alkynyl, aryl, or heteroaryl moieties may be linked to the remainder of the molecule via an alkylene bond. Under these circumstances, the substituent will be referred to as alkenylalkyl, alkynylalkyl, arylalkyl, or heteroarylalkyl, indicating that the alkylene moiety is between the alkenyl, alkynyl, aryl, or heteroaryl and the molecule to which the alkenyl, alkynyl, aryl, or heteroaryl is bonded.

The term "halogen" includes bromine, fluorine, chlorine and iodine.

The term "heterocyclic ring" refers to a 4 to 8 membered aromatic or non-aromatic ring containing 3 to 7 carbon atoms and at least one N, O, or S atom. Examples are piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidine and tetrahydrofuranyl, as well as exemplary groups given above for the term "heteroaryl".

When the ring system is optionally substituted, suitable substituents are selected from the group consisting of alkyl, alkenyl, alkynyl, or an additional ring, where each is optionally further substituted. Optional substituents in any group including those listed above include halo, nitro, cyano, OR, -SR, -NR ₂ , -OCOR, -NRCOR, -COOR, -CONR ₂ , -SOR, -SO ₂ R, -SONR ₂ , -SO ₂ NR ₂ where each R is independently alkyl, alkenyl, alkynyl, aryl or heteroaryl, or two R groups taken together with the atoms to which they are attached form a ring.

Preferably -L ¹ - of formula (IV) is substituted with one component -L ² -Z or L ² -Z'.

An additional preferred option -L ¹ - is disclosed in WO2013/036857A1, which is incorporated herein in its entirety by reference. Accordingly, the preferred component -L ¹ - has the formula (V):

(V),

(V)

where

the dotted line shows the attachment to -D, which is

PTH component, and where the accession is through the amine functional group of the component -D;

-R ¹ is selected from the group consisting of optionally substituted C ₁ -C ₆ linear, branched, or cyclic alkyl; optionally substituted aryl; optionally substituted heteroaryl; alkoxy; and NR ⁵ ₂ ;

-R ² is selected from the group consisting of -H; optionally substituted C ₁ -C ₆ alkyl; optionally substituted aryl; and optionally substituted heteroaryl;

-R ³ is selected from the group consisting of -H; optionally substituted C ₁ -C ₆ alkyl; optionally substituted aryl; and optionally substituted heteroaryl;

-R ⁴ is selected from the group consisting of -H; optionally substituted C ₁ -C ₆ alkyl; optionally substituted aryl; and optionally substituted heteroaryl;

each -R ⁵ is independently selected from the group consisting of -H; optionally substituted C ₁ -C ₆ alkyl; optionally substituted aryl; and optionally substituted heteroaryl; or taken together two -R ⁵ may be cycloalkyl or cycloheteroalkyl;

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Only in the context of formula (V) the terms used have the following meanings:

"Alkyl", "alkenyl" and "alkynyl" include linear, branched or cyclic hydrocarbon groups having 1 to 8 carbon atoms or 1 to 6 carbon atoms or 1 to 4 carbon atoms, where alkyl is a saturated hydrocarbon, alkenyl includes one or more carbon-carbon double bonds, and alkynyl includes one or more carbon-carbon triple bonds. Unless otherwise indicated, they contain 1-6 carbon atoms.

"Aryl" includes aromatic hydrocarbon groups having 6 to 18 carbon atoms, preferably 6 to 10 carbon atoms, including groups such as phenyl, naphthyl and anthracene. "Heteroaryl" includes aromatic rings containing 3-15 carbon atoms containing at least one N, O or S atom, preferably 3-7 carbon atoms containing at least one N, O or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isooxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl and the like.

The term “substituted” means an alkyl, alkenyl, alkynyl, aryl, or heteroaryl group having one or more substituent groups instead of one or more hydrogen atoms. Substituents may generally be selected from halogen, including F, Cl, Br and I; lower alkyl, including linear, branched and cyclic; lower haloalkyl including fluoroalkyl, chloroalkyl, bromoalkyl and iodoalkyl; HE; lower alkoxy, including linear, branched and cyclic; SH; lower alkylthio, including linear, branched and cyclic; amino, alkylamino, dialkylamino, silyl, including alkylsilyl, alkoxysilyl and arylsilyl; nitro; cyano; carbonyl; carboxylic acid, carboxylic acid ester, carboxylic amide, aminocarbonyl; aminoacyl; carbamate; urea; thiocarbamate; thiourea; ketone; sulfone; sulfonamide; aryl, including phenyl, naphthyl and anthracenyl; heteroaryls, including 5-membered heteroaryls, including pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole, and tetrazole, 6-membered heteroaryls, including pyridine, pyrimidine, pyrazine, and fused heteroaryls including benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzisoxazole and benzisothiazole.

Preferably -L ¹ - of formula (V) is substituted with one component -L ² -Z or L ² -Z'.

Another preferred option -L ¹ - disclosed in US7585837B2, which is incorporated into this application in its entirety by reference. Accordingly, the preferred component -L ¹ - has the formula (VI):

where

the dotted line shows attachment to -D, which is the PTH moiety, and where the attachment is through the amine functional group of the -D moiety;

R ¹ and R ² are independently selected from the group consisting of hydrogen, alkyl, alkoxy, alkoxyalkyl, aryl, alkaryl, aralkyl, halogen, nitro, -SO ₃ H, -SO ₂ NHR ⁵ , amino, ammonium, carboxyl, PO ₃ H ₂ and OPO ₃ H ₂ ;

R ³ , R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, alkyl and aryl;

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Suitable substituents for formula (VI) are alkyl (such as C _1-6 alkyl), alkenyl (such as C _2-6 alkenyl), alkynyl (such as C _2-6 alkynyl), aryl (such as phenyl), heteroalkyl , heteroalkenyl, heteroalkynyl, heteroaryl (such as an aromatic 4-7 membered heterocycle) or halogen.

Only in the context of formula (VI) the terms used have the following meanings:

The terms "alkyl", "alkoxy", "alkoxyalkyl", "aryl", "alkaryl" and "aralkyl" mean alkyl radicals containing 1-8, preferably 1-4 carbon atoms, such as methyl, ethyl, propyl, isopropyl and butyl, and aryl radicals containing 6-10 carbon atoms, such as phenyl and naphthyl. The term "halogen" includes bromine, fluorine, chlorine and iodine.

Preferably -L ¹ - of formula (VI) has as a substituent one component -L ² -Z or L ² -Z'.

Another preferred option -L ¹ - disclosed in WO2002/089789A1, which is incorporated into this application in its entirety by reference. Accordingly, the preferred component -L ¹ - has the formula (VII):

where

the dotted line shows attachment to -D, which is the PTH moiety, and where the attachment is through the amine functional group of the -D moiety;

L ₁ is a bifunctional linking group,

Y ₁ and Y ₂ are independently O, S or NR ⁷ ;

R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are independently selected from the group consisting of hydrogen, C _1-6 alkyl, C _3-12 branched alkyl, C _3-8 cycloalkyl, C _1-6 substituted alkyls, C _3-8 substituted cycloalkyls, aryls, substituted aryls, aralkyls, C _1-6 heteroalkyls, substituted C _1-6 heteroalkyls, C _1-6 alkoxy, phenoxy and C _1-6 heteroalkoxy;

Ar is a moiety which, when included in formula (VII), forms a multi-substituted aromatic hydrocarbon or a multi-substituted heterocyclic group;

X is a chemical bond or moiety that is actively transported into the target cell, a hydrophobic moiety, or a combination thereof,

y is 0 or 1;

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Only in the context of formula (VII) the terms used have the following meanings:

The term “alkyl” is to be understood to include, for example, linear, branched, substituted C _1-12 alkyls, including alkoxy, C _3-8 cycloalkyls or substituted cycloalkyls, etc.

The term "substitution", as should be understood, includes the addition or replacement of one or more atoms contained in the functional group, or compounds with one or more different atoms.

Substituted alkyls include carboxyalkyls, aminoalkyls, dialkylamines, hydroxyalkyls and mercaptoalkyls; substituted cycloalkyls include moieties such as 4-chlorocyclohexyl; aryls include constituents such as naphthyl; substituted aryls include moieties such as 3-bromo-phenyl; aralkyls include moieties such as toluyl; heteroalkyls include moieties such as ethylthiophene; substituted heteroalkyls include moieties such as 3-methoxythiophene; alkoxy includes moieties such as methoxy; and phenoxy includes moieties such as 3-nitrophenoxy. halo is to be understood to include fluorine, chlorine, iodine and bromine.

Preferably -L ¹ - of formula (VII) has as a substituent one component -L ² -Z or L ² -Z'.

In another preferred embodiment of the present invention -L ¹ - contains a substructure of formula (VIII)

(VIII),

(VIII)

where

the dotted line marked with an asterisk represents the attachment to the nitrogen atom of the -D moiety, which is the PTH moiety, by forming an amide bond;

unmarked dotted lines show attachment to the remainder of -L ¹ -; and

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Preferably -L ¹ - of formula (VIII) is substituted with one component -L ² -Z or L ² -Z'.

In one embodiment of the present invention, -L ¹ - of formula (VIII) is not further substituted.

In another preferred embodiment of the present invention -L ¹ - contains a substructure of formula (IX)

(IX),

(IX)

where

the dotted line marked with an asterisk represents the attachment to the nitrogen atom of the -D moiety, which is the PTH moiety, by forming a carbamate bond;

unmarked dotted lines show attachment to the remainder of -L ¹ -; and

where -L ¹ - is substituted with -L ² -Z or L ² -Z', and where -L ¹ - is optionally further substituted;

where

-L ² - represents a simple chemical bond or spacer;

-Z is a water-soluble carrier; and

-Z' is a water-insoluble carrier.

Preferably -L ¹ - of formula (IX) is substituted with one component -L ² -Z or L ² -Z'.

In one embodiment of the present invention, -L ¹ - of formula (IX) is not further substituted.

In prodrugs for use according to the present invention, -L ² - is a chemical binder or spacer component.

In one embodiment of the present invention, -L ² - is a chemical bond.

In another embodiment of the present invention -L ² - represents the spacer component.

When -L ² is other than a single chemical bond, -L ² - is preferably selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O) N(R ^y1 )-, -S(O) ₂ N(R ^y1 )-, -S(O)N(R ^y1 )-, -S(O) ₂ -, -S(O)-, -N( R ^y1 )S(O) ₂ N(R ^y1a )-, -S-, -N(R ^y1 )-, -OC(OR ^y1 )(R ^y1a )-, -N(R ^y1 )C(O)N (R ^y1a )-, -OC(O)N(R ^y1 )-, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where -T-, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally substituted with one or more -R ^y2 that are the same or different, and where C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ^y3 )-, -S(O) ₂ N(R ^y3 )-, -S(O)N(R ^y3 )-, -S(O) ₂ -, -S(O)-, -N(R ^y3 ) S(O) ₂ N(R ^y3a )-, -S-, -N(R ^y3 )-, -OC(OR ^y3 )(R ^y3a )-, -N(R ^y3 )C(O)N(R ^y3a )-, and OC(O)N(R ^y3 )-;

-R ^y1 and R ^{y1a are} independently selected from the group consisting of H, -T, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where -T, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally substituted with one or more -R ^y2 that are the same or different, and where C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl is optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ^y4 )-, -S(O) ₂ N(R ^y4 )-, -S(O)N(R ^y4 )-, -S(O) ₂ -, -S(O)-, -N(R ^y4 )S (O) ₂ N(R ^y4a )-, -S-, -N(R ^y4 )-, -OC(OR ^y4 )(R ^y4a )-, -N(R ^y4 )C(O)N(R ^y4a ) -, and OC(O)N(R ^y4 )-;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10 cycloalkyl, _3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl; where each T is independently optionally substituted with one or more -R ^y2 that are the same or different;

each -R ^y2 is independently selected from the group consisting of halogen, -CN, oxo (=O), -COOR ^y5 , -OR ^y5 , -C(O)R ^y5 , -C(O)N(R ^y5 R ^y5a ) , -S(O) ₂ N(R ^y5 R ^y5a ), -S(O)N(R ^y5 R ^y5a ), -S(O) ₂ R ^y5 , -S(O)R ^y5 , -N(R ^y5 )S(O) ₂ N(R ^y5a R ^y5b ), -SR ^y5 , -N(R ^y5 R ^y5a ), -NO ₂ , -OC(O)R ^y5 , -N(R ^y5 )C(O)R ^y5a , -N(R ^y5 )S(O) ₂ R ^y5a , -N(R ^y5 )S(O)R ^y5a , -N(R ^y5 )C(O)OR ^y5a , -N(R ^y5 )C( O)N(R ^y5a R ^y5b ), -OC(O)N(R ^y5 R ^y5a ), and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogens that are the same or different; and

-R ^y3 , -R ^y3a , -R ^y4 , -R ^y4a , -R ^y5 , -R ^y5a and R ^{y5b are each} independently selected from the group consisting of -H and C _1-6 alkyl, where C _1-6 alkyl optionally substituted with one or more halogens which are the same or different.

When -L ² - is other than a single chemical bond, -L ² - is even more preferably selected from -T-, -C(O)O-, -O-, -C(O)-, -C(O)N( R ^y1 )-, -S(O) ₂ N(R ^y1 )-, -S(O)N(R ^y1 )-, -S(O) ₂ -, -S(O)-, -N(R ^y1 )S(O) ₂ N(R ^y1a )-, -S-, -N(R ^y1 )-, -OC(OR ^y1 )(R ^y1a )-, -N(R ^y1 )C(O)N(R ^y1a )-, -OC(O)N(R ^y1 )-, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where -T-, C _1-20 alkyl, C _2-20 alkenyl, and C _2-20 alkynyl are optionally substituted with one or more -R ^y2 that are the same or different, and where C _1-20 alkyl, C _2-20 alkenyl and C _2-20 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N( R ^y3 )-, -S(O) ₂ N(R ^y3 )-, -S(O)N(R ^y3 )-, -S(O) ₂ -, -S(O)-, -N(R ^y3 )S(O) ₂ N(R ^y3a )-, -S-, -N(R ^y3 )-, -OC(OR ^y3 )(R ^y3a )-, -N(R ^y3 )C(O)N(R ^y3a )-, and OC(O)N(R ^y3 )-;

-R ^y1 and R ^{y1a are} independently selected from the group consisting of H, -T, C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl; where -T, C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl are optionally substituted with one or more -R ^y2 that are the same or different, and where C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl is optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ^y4 )-, -S(O) ₂ N(R ^y4 )-, -S(O)N(R ^y4 )-, -S(O) ₂ -, -S(O)-, -N(R ^y4 )S (O) ₂ N(R ^y4a )-, -S-, -N(R ^y4 )-, -OC(OR ^y4 )(R ^y4a )-, -N(R ^y4 )C(O)N(R ^y4a ) -, and OC(O)N(R ^y4 )-;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10 cycloalkyl, _3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl; where each T is independently optionally substituted with one or more -R ^y2 that are the same or different;

-R ^y2 is selected from the group consisting of halogen, -CN, oxo (=O), -COOR ^y5 , -OR ^y5 , -C(O)R ^y5 , -C(O)N(R ^y5 R ^y5a ), - S(O) ₂ N(R ^y5 R ^y5a ), -S(O)N(R ^y5 R ^y5a ), -S(O) ₂ R ^y5 , -S(O)R ^y5 , -N(R ^y5 )S (O) ₂ N(R ^y5a R ^y5b ), -SR ^y5 , -N(R ^y5 R ^y5a ), -NO ₂ , -OC(O)R ^y5 , -N(R ^y5 )C(O)R ^y5a , -N(R ^y5 )S(O) ₂ R ^y5a , -N(R ^y5 )S(O)R ^y5a , -N(R ^y5 )C(O)OR ^y5a , -N(R ^y5 )C(O) N(R ^y5a R ^y5b ), -OC(O)N(R ^y5 R ^y5a ), and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogens that are the same or different; and

-R ^y3 , -R ^y3a , -R ^y4 , -R ^y4a , -R ^y5 , -R ^y5a and R ^{y5b are} each independently selected from the group consisting of H and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogens which are the same or different.

When -L ² - is other than a single chemical bond, -L ² - is even more preferably selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C( O)N(R ^y1 )-, -S(O) ₂ N(R ^y1 )-, -S(O)N(R ^y1 )-, -S(O) ₂ -, -S(O)-, - N(R ^y1 )S(O) ₂ N(R ^y1a )-, -S-, -N(R ^y1 )-, -OC(OR ^y1 )(R ^y1a )-, -N(R ^y1 )C(O )N(R ^y1a )-, -OC(O)N(R ^y1 )-, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where -T-, C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally substituted with one or more -R ^y2 that are the same or different, and where C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ^y3 )-, -S(O) ₂ N(R ^y3 )-, -S(O)N(R ^y3 )-, -S(O) ₂ -, -S(O)-, -N(R ^y3 ) S(O) ₂ N(R ^y3a )-, -S-, -N(R ^y3 )-, -OC(OR ^y3 )(R ^y3a )-, -N(R ^y3 )C(O)N(R ^y3a )-, and OC(O)N(R ^y3 )-;

-R ^y1 and R ^y1a are independently selected from the group consisting of H, -T, C _1-10 alkyl, C _2-10 alkenyl, and C _2-10 alkynyl;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10 cycloalkyl, _3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl;

each -R ^y2 is independently selected from the group consisting of halo and C _1-6 alkyl; and

-R ^y3 , -R ^y3a , -R ^y4 , -R ^y4a , -R ^y5 , -R ^y5a and R ^{y5b are} each independently selected from the group consisting of H and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogens which are the same or different.

Even more preferably, -L ² - is a C _1-20 alkyl chain which is optionally interrupted by one or more groups independently selected from -O-, -T- and -C(O)N(R ^y1 )-; and where the C _1-20 alkyl chain is optionally substituted with one or more groups independently selected from -OH, -T and C(O)N(R ^y6 R ^y6a ); where -R ^y1 , -R ^y6 , -R ^y6a are independently selected from the group consisting of H and C _1-4 alkyl, and where T is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C _3-10 cycloalkyl, 3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl.

Preferably, -L ² - has a molecular weight in the range from 14 g/mol to 750 g/mol.

Preferably, -L ² - contains a moiety selected from

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, и

;

,

, and

;

where

the dotted line denotes attachment to the rest of -L ² -, -L ¹ -, -Z and/or -Z', respectively; and

-R and R ^a are independently selected from the group consisting of H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

In one preferred embodiment of the present invention - L ² - has a chain length of 1 to 20 atoms.

As used in this application, the term "chain length" in relation to the -L ² - component refers to the number of -L ² - atoms present in the shortest connection between -L ¹ - and Z.

Preferably, -L ² - has the formula (i)

(i),

(i)

where

dashed line marked with an asterisk means attachment to -L ¹ -;

an unmarked dotted line denotes attachment to -Z or Z';

n is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18; and

where the component of formula (i) is optionally additionally substituted.

Preferably, n of formula (i) is selected from the group consisting of 3, 4, 5, 6, 7, 8, and 9. Even more preferably, n of formula (i) is 4, 5, 6, or 7. In one embodiment, of the present invention, n of formula (i) is 4. In another embodiment of the present invention, n of formula (i) is 5. In another embodiment of the present invention, n of formula (i) is 6.

(IIca-i),In another preferred embodiment of the present invention, the component -L ¹ -L ² - is selected from the group consisting of

(IIca-i),

(IIca-ii) и

(IIca-ii) and

(IIca-iii);

(IIca-iii);

where

the unmarked dotted line denotes the attachment to the nitrogen atom of the D moiety, which is the PTH moiety, by forming an amide bond; and

a dotted line marked with an asterisk means attached to -Z or -Z'.

In one preferred embodiment of the present invention, the component -L ¹ -L ² - is selected from the group consisting of

(IIcb-i),

(IIcb-i),

(IIcb-ii) и

(IIcb-ii) and

(IIcb-iii);

(IIcb-iii);

the unmarked dotted line denotes the attachment to the nitrogen atom of the D moiety, which is the PTH moiety, by forming an amide bond; and

a dotted line marked with an asterisk means attached to -Z or -Z'.

In a preferred embodiment of the present invention, the component -L ¹ -L ² - has the formula (IIca-ii).

Preferably, the controlled release PTH compound of the present invention has the formula (Ia) with x=1.

The -Z carrier contains C _8-24 alkyl or polymer. Preferably, -Z contains a polymer, preferably a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphoylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy)polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly (dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl- oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-glycolic acid copolymers), poly(methacrylamides), poly (methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinylmethyl ethers), poly(vinyl pyrrolidones), silicones , celluloses, carbomethylcelluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other polymers based on carbohydrates, xylans and their copolymers.

Preferably, -Z has a molecular weight in the range of 5 to 200 kDa. Even more preferably, -Z has a molecular weight in the range of 8 to 100 kDa, even more preferably in the range of 10 to 80 kDa, even more preferably 12 to 60, even more preferably 15 to 40, and most preferably -Z has molecular weight of about 20 kDa. In another equally preferred embodiment of the present invention, -Z has a molecular weight of about 40 kD.

In one embodiment of the present invention, such a water-soluble carrier -Z contains a protein. Preferred proteins are selected from the group consisting of carboxyl-terminal human chorionic gonadotropin polypeptide as described in US 2012/0035101 A1, which are incorporated herein by reference; albumin; XTEN as described in WO 2011123813 A2, which is incorporated herein by reference; proline/alanine random coil sequences as described in WO 2011/144756 A1, which is incorporated herein by reference; proline/alanine/serine sequences as described in WO 2008/155134 A1 and WO 2013/024049 A1, which are incorporated herein by reference; with Fc fusion proteins.

In one embodiment of the present invention, -Z is a polysarcosine.

In another preferred embodiment of the present invention, -Z contains poly(N-methylglycine).

In a particularly preferred embodiment of the present invention, the -Z contains a protein component with a random helix structure.

In one preferred embodiment of the present invention, -Z contains a single protein moiety with a random helix structure.

In another preferred embodiment of the present invention, -Z contains two protein components with a random helix structure.

In another preferred embodiment of the present invention, -Z contains three protein components with a random helix structure.

In another preferred embodiment of the present invention, -Z contains four protein components with a random helix structure.

In another preferred embodiment of the present invention, -Z contains five protein components with a random helix structure.

In another preferred embodiment of the present invention, -Z contains six protein components with a random helix structure.

In another preferred embodiment of the present invention, -Z contains seven protein components with a random helix structure.

In another preferred embodiment of the present invention, -Z contains eight protein components with a random helix structure.

Preferably, the random coil protein moiety contains at least 25 amino acid residues and at most 2000 amino acid residues. Even more preferably, the random coil protein moiety contains at least 30 amino acid residues and at most 1500 amino acid residues. Even more preferably, the random coil protein component contains at least 50 amino acid residues and at most 500 amino acid residues.

In a preferred embodiment of the present invention, -Z comprises a protein moiety with a random helix structure, wherein at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said random coil protein component are selected from alanine and proline. Even more preferably, at least 10%, but less than 75%, preferably less than 65%, of the total number of amino acid residues of the random coil protein component are proline residues. Preferably, the random coil protein component is as described in WO 2011/144756 A1, which is incorporated herein in its entirety by reference. Even more preferably, Z contains at least one moiety selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO :6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14 , SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:51 and SEQ ID NO:61 as disclosed in WO2011/144756, which is incorporated herein by reference. A moiety containing such a protein with a random helix structure containing alanine and proline, which is referred to as a “PA” or “PA moiety”.

Accordingly, -Z contains a PA component.

In an equally preferred embodiment of the present invention, -Z comprises a random coil protein moiety in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95 %, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids that form the specified component of the protein with a random helix structure, selected from alanine, serine and proline. Even more preferably, at least 4% but less than 40% of the total amino acid residues of the random coil protein component are proline residues. Preferably, the random coil protein component is described in WO 2008/155134 A1, which is incorporated herein in its entirety by reference. Even more preferably, Z contains at least one moiety selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO :12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28 , SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 and SEQ ID NO:56 as described in WO 2008/155134 A1, which is incorporated herein by reference. A moiety containing a protein with a random helix structure containing alanine, serine and proline is referred to as a “PAS” or “PAS moiety”.

Accordingly, -Z contains a PAS component.

In an equally preferred embodiment of the present invention, -Z comprises a random coil protein moiety in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95 %, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids that form the specified component of the protein with a reverse helix structure, selected alanine, glycine and proline. A moiety containing a protein with a random helix structure containing alanine, glycine and proline is referred to as "PAG" or "PAG moiety".

Accordingly, -Z contains a PAG component.

In an equally preferred embodiment of the present invention, -Z comprises a random coil protein moiety in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95 %, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids that form the specified component of the protein with a reverse helix structure, is selected from proline and glycine. A moiety containing a protein with a random helix structure containing proline and glycine is referred to as a “PG” or “PG moiety”.

Preferably, such a PG moiety contains a moiety of formula (a-0)

[(Gly) _p -Pro-(Gly) _q ] _r (a-0);

where

p is selected from the group consisting of 0, 1, 2, 3, 4 and 5;

q is selected from the group consisting of 0, 1, 2, 3, 4 and 5;

r is an integer in the range and inclusive of 10 to 1000;

provided that at least one of p and q is at least 1;

Preferably, p of formula (a-0) is selected from the group consisting of 1, 2 and 3.

Preferably, q of formula (a-0) is selected from 0, 1 and 2.

Even more preferably, the PG moiety contains the sequence SEQ ID: NO 122: GGPGGPGPGGPGGPGPGGPG

Even more preferably, the PG moiety contains the sequence SEQ ID: NO 97 of formula (a-0-a)

(GGPGGPGPGGPGGPGPGGPG) _v (a-0-a),

where

v is an integer in the range and inclusive of 1 to 50.

It is understood that the sequence of formula (a-0-a) contains v repetitions of the sequence SEQ ID NO:122.

Accordingly, -Z contains a PG component.

In an equally preferred embodiment of the present invention, -Z comprises a random coil protein moiety in which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95 %, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said reverse helix protein are selected from alanine, glycine, serine, threonine, glutamate and proline. Preferably, the random coil protein component is as described in WO 2010/091122 A1, which is incorporated herein by reference. Even more preferably -Z contains at least one member selected from the group consisting of SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184; SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO: 201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219, SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:759, SEQ ID NO:760, SEQ ID NO:761, SEQ ID NO:762, SEQ ID NO: 763, SEQ ID NO:764, SEQ ID NO:765, SEQ ID NO:766, SEQ ID NO:767, SEQ ID NO:768, SEQ ID NO:769, SEQ ID NO:770, SEQ ID NO:771, SEQ ID NO:772, SEQ ID NO:773, SEQ ID NO:774, SEQ ID NO:775, SEQ ID NO:776, SEQ ID NO:777, SEQ ID NO:778, SEQ ID NO:779, SEQ ID NO:1715, SEQ ID NO:1716, SEQ ID NO:1718, SEQ ID NO:1719, SEQ ID NO:1720, SEQ ID NO:1721 and SEQ ID NO:1722 as described in WO2 010/091122A1, which is incorporated into this application by reference. A moiety containing a protein with a random helix structure containing alanine, glycine, serine, threonine, glutamate and proline is referred to as "XTEN" or "XTEN moiety" along with its designation in WO 2010/091122 A1.

Accordingly, -Z contains the XTEN component.

In another preferred embodiment of the present invention, Z contains a fatty acid derivative. Preferred fatty acid derivatives are those described in WO 2005/027978 A2 and WO 2014/060512 A1, which are incorporated herein by reference.

In another preferred embodiment of the present invention, Z is a polymer based on hyaluronic acid.

In one embodiment of the present invention, Z is a carrier as disclosed in WO 2012/02047 A1, which is incorporated herein by reference.

In another embodiment of the present invention, Z is a carrier as disclosed in WO 2013/024048 A1, which is incorporated herein by reference.

In another preferred embodiment of the present invention, -Z is a PEG-based polymer, such as a linear, branched or multi-branched PEG-based polymer.

In one embodiment of the present invention, -Z is a linear PEG-based polymer.

In another embodiment of the present invention, -Z is a multi-armed PEG polymer. Preferably, -Z is a multi-arm PEG polymer having at least 4 PEG arms.

Preferably, such PEG-based polymer with multiple -Z arms is connected to a plurality of -L ² -L ¹ -D moieties, where each -L ² -L ¹ -D moiety is preferably connected to the end of the leg, preferably the end of the leg. Preferably such a PEG-based polymer with multiple branches -Z is connected with 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 constituents -L ² -L ¹ - D. Even more preferably, such a multi-Z arm PEG polymer is coupled to 2, 3, 4, 6 or 8 -L ² -L ¹ - D moieties. , 4 or 6 moieties -L ² -L ¹ -D, even more preferably such a PEG-based polymer with multiple -Z branches is coupled to 4 or 6 moieties -L ² -L ¹ -D, and most preferably such a PEG-based polymer with many branches -Z is connected to 4 components -L ² -L ¹ -D.

Preferably such multi-armed PEG-based polymer Z is a multi-armed PEG derivative, as for example detailed for the products listed in JenKem Technology, USA (available at http://www.jenkemusa.com/Pages/PEGProducts on December 18 2014), such as a 4-arm PEG derivative, in particular a 4-arm PEG derivative containing a pentaerythritol core, an 8-arm PEG derivative containing a hexaglycerol core, and an 8-arm PEG derivative containing a tripentaerythritol core . More preferably, the water-soluble PEG-based -Z carrier contains a moiety selected from:

PEG-amine with 4 branches containing a core of pentaerythritol:

with n in the range from 20 to 500;

PEG-amine with eight branches containing a core of hexaglycerol:

with n in the range from 20 to 500; and

R=core structure of hexaglycerol or tripentaerythritol; and

PEG-amine with six branches containing a core of sorbitol or dipentaerythritol:

with n in the range from 20 to 500; and

R=containing a core of sorbitol or dipentaerythritol;

and where dotted lines represent attachment to the rest of the PTH prodrug.

In a preferred embodiment of the present invention, -Z is a branched PEG-based polymer. In one embodiment of the present invention, -Z is a branched PEG-based polymer having one, two, three, four, five, or six branch points. Preferably, -Z is a branched PEG-based polymer having one, two or three branch points. In one embodiment of the present invention, -Z is a branched PEG-based polymer having a single branch point. In another embodiment of the present invention, -Z is a branched PEG-based polymer having two branching points. In another embodiment of the present invention, -Z is a branched PEG-based polymer having three branching points.

The branching point is preferably selected from the group consisting of -N<, -CH< and >C<.

Preferably, such a branched -Z moiety has a molecular weight of at least 10 kDa.

In one embodiment of the present invention, such a branched -Z moiety has a molecular weight in the range and including 10 kDa - 500 kDa, more preferably in the range and including 10 kDa - 250 kDa, even more preferably in the range and including 10 kDa - 150 kDa, even more preferably in the range and including 12 kDa - 100 kDa and most preferably in the range and including 15 kDa - 80 kDa.

Preferably, such a branched -Z moiety has a molecular weight in the range and including 10 kDa - 80 kDa. In one embodiment of the present invention, the molecular weight is about 10 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 20 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 30 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 40 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 50 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 60 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 70 kDa. In another embodiment of the present invention, the molecular weight of such a branched -Z moiety is about 80 kDa. Most preferably, such a branched -Z moiety has a molecular weight of about 40 kDa.

Preferably, -Z or Z' contains a component

.

.

In an equally preferred embodiment of the present invention, Z contains an amide bond.

Preferably -Z contains a component of formula (a)

(a),

(a)

where

the dotted line shows attachment to -L ² - or to the remainder of -Z;

BP ^a is a branch point selected from the group consisting of -N<, -CR<and>C<;

-R is selected from the group consisting of -H and C _1-6 alkyl;

a is 0 if BP ^a is -N< or CR<, and n is 1 if BP ^a is >C<;

-S ^a -, -S ^a' -, -S ^a'' - and S ^a''' - independently of each other represent a chemical bond or are selected from the group consisting of C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl; where C _1-50 alkyl, C _2-50 alkenyl and C _2-50 alkynyl are optionally substituted with one or more substituents -R ¹ that are the same or different, and where C _1-50 alkyl, C _2-50 alkenyl and C _{2 -50} alkynyl optionally interrupted by one or more groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N(R ² )- , -S(O) ₂ N(R ² )-, -S(O)N(R ² )-, -S(O) ₂ -, -S(O)-, -N(R ² )S(O ) ₂ N(R ^2a )-, -S-, -N(R ² )-, -OC(OR ² )(R ^2a )-, -N(R ² )C(O)N(R ^2a )- and -OC(O)N(R ² )-;

each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10 cycloalkyl, _3-10 membered heterocyclyl, 8-11 membered heterobicyclyl, 8-30 membered carbopolycyclyl, and 8-30 membered heteropolycyclyl; where each -T- is independently optionally substituted with one or more substituents -R ¹ that are the same or different;

each -R ¹ is independently selected from the group consisting of halogen, -CN, oxo (=O), -COOR ³ , -OR ³ , -C(O)R ³ , -C(O)N(R ³ R ^3a ) , -S(O) ₂ N(R ³ R ^3a ), -S(O)N(R ³ R ^3a ), -S(O) ₂ R ³ , -S(O)R ³ , -N(R ³ )S(O) ₂ N(R ^3a R ^3b ), -SR ³ , -N(R ³ R ^3a ), -NO ₂ , -OC(O)R ³ , -N(R ³ )C(O)R ^3a , -N(R ³ )S(O) ₂ R ^3a , -N(R ³ )S(O)R ^3a , -N(R ³ )C(O)OR ^3a , -N(R ³ )C( O)N(R ^3a R ^3b ), -OC(O)N(R ³ R ^3a ) and C _1-6 alkyl; where C _1-6 alkyl is optionally substituted with one or more halogen substituents that are the same or different;

each -R ² , -R ^2a , -R ³ , -R ^3a and R ^3b are independently selected from the group consisting of -H and C _1-6 alkyl, where C _1-6 alkyl is optionally substituted with one or more halogen substituents which are are the same or different; and

-P ^a' , -P ^a'' and P ^a''' are independently a polymeric moiety.

In one embodiment of the present invention, BP ^a of formula (a) is -N<.

In another embodiment of the present invention, BP ^a of formula (a) is >C<.

In a preferred embodiment of the present invention, BP ^a of formula (a) is -CR<. Preferably, -R is -H. Accordingly, a of formula (a) is equal to 0.

In one embodiment of the present invention, the -S ^a - of formula (a) is a chemical bond.

In another embodiment of the present invention, -S ^a - of formula (a) is selected from the group consisting of C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl, where C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of -T-, -C(O)O-, -O-, -C(O)-, -C(O)N( R ⁴ )-, -S(O) ₂ N(R ⁴ )-, -S(O)N(R ⁴ )-, -S(O) ₂ -, -S(O)-, -N(R ⁴ )S(O) ₂ N(R ^4a )-, -S-, -N(R ⁴ )-, -OC(OR ⁴ )(R ^4a )-, -N(R ⁴ )C(O)N(R ^4a )-, and OC(O)N(R ⁴ )-; where -T- is a 3-10 membered heterocyclyl; and R ⁴ and R ^4a are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl.

Preferably -S ^a - of formula (a) is selected from the group consisting of C _1-10 alkyl, which is interrupted by one or more chemical groups selected from the group consisting of -T-, -C(O)N(R ⁴ )- and O-.

In one embodiment of the present invention, the -S ^a' - of formula (a) is a chemical bond.

In another embodiment of the present invention, -S ^a' - formula (a) is selected from the group consisting of C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl, where C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of -C(O)O-, -O-, -C(O)-, -C(O)N(R ⁴ ) -, -S(O) ₂ N(R ⁴ )-, -S(O)N(R ⁴ )-, -S(O) ₂ -, -S(O)-, -N(R ⁴ )S( O) ₂ N(R ^4a )-, -S-, -N(R ⁴ )-, -OC(OR ⁴ )(R ^4a )-, -N(R ⁴ )C(O)N(R ^4a )- , and OC(O)N(R ⁴ )-; where -R ⁴ and R ^4a are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Preferably -S ^a' - of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and C(O)N(R ⁴ )-.

In one embodiment of the present invention -S ^a'' - of formula (a) is a chemical bond.

In another embodiment of the present invention -S ^a'' - formula (a) is selected from the group consisting of C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl, where C _1-10 alkyl, C _{2- 10} alkenyl and C _2-10 alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of -C(O)O-, -O-, -C(O)-, -C(O)N(R ⁴ )-, -S(O) ₂ N(R ⁴ )-, -S(O)N(R ⁴ )-,-S(O) ₂ -, -S(O)-, -N(R ⁴ )S (O) ₂ N(R ^4a )-, -S-, -N(R ⁴ )-, -OC(OR ⁴ )(R ^4a )-, -N(R ⁴ )C(O)N(R ^4a ) -, and OC(O)N(R ⁴ )-; where -R ⁴ and R ^4a are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Preferably -S ^a ''- of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O)- and C(O)N(R ⁴ )-.

In one embodiment of the present invention, -S ^a''' - of formula (a) is a chemical bond.

In another embodiment of the present invention -S ^a''' - of formula (a) is selected from the group consisting of C _1-10 alkyl, C _2-10 alkenyl and C _2-10 alkynyl, where C _1-10 alkyl, C _{2 -10} alkenyl and C _2-10 alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of -C(O)O-, -O-, -C(O)-, -C(O)N(R ⁴ )-, -S(O) ₂ N(R ⁴ )-, -S(O)N(R ⁴ )-,-S(O) ₂ -, -S(O)-, -N(R ⁴ ) S(O) ₂ N(R ^4a )-, -S-, -N(R ⁴ )-, -OC(OR ⁴ )(R ^4a )-, -N(R ⁴ )C(O)N(R ^4a )-, and OC(O)N(R ⁴ )-; where -R ⁴ and R ^4a are independently selected from the group consisting of H, methyl, ethyl, propyl and butyl. Preferably -S ^a''' - formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of -O-, -C(O) - and C(O)N(R ⁴ )-.

Preferably, -P ^a' , -P ^a'' and P ^a''' of formula (a) independently contain a polymer selected from the group consisting of 2-methacryloyl-oxyethylphosphoylcholines, poly(acrylic acids), poly(acrylates), poly (acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly (caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazolines), poly(iminocarbonates), poly(lactic acids), poly(copolymers of lactic and glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(or ganophosphazenes), poly(orthoesters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinylmethyl ethers), poly(vinyl pyrrolidones) ), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers , xylans and their copolymers.

More preferably, -P ^a' , -P ^a'' and P ^a''' of formula (a) independently contain a PEG-based moiety. Even more preferably, -P ^a' , -P ^a'' and P ^a''' of formula (a) independently contain a PEG-based moiety containing at least 20% PEG, even more preferably at least 30%, even more preferably at least 40% PEG, even more preferably at least 50% PEG, even more preferably at least 60% PEG, even more preferably at least 70% PEG, even more preferably at least 80% PEG, and most preferably at least 90% PEG.

Preferably, -P ^a' , -P ^a'' and P ^a''' of formula (a) independently have a molecular weight in the range and including 5 kDa - 50 kDa, more preferably have a molecular weight in the range and including 5 kDa - 40 kDa , even more preferably in the range and including 7.5 kDa - 35 kDa, even more preferably in the range of and 7.5 - 30 kDa, even more preferably in the range and including 10 - 30 kDa.

In one embodiment of the present invention -P ^a' , -P ^a'' and P ^a''' of formula (a) have a molecular weight of about 5 kDa.

In another embodiment of the present invention -P ^a' , -P ^a'' and P ^a''' of formula (a) have a molecular weight of about 7.5 kDa.

In another embodiment of the present invention -P ^a' , -P ^a'' and P ^a''' of formula (a) have a molecular weight of about 10 kDa.

In another embodiment of the present invention -P ^a' , -P ^a'' and P ^a''' of formula (a) have a molecular weight of about 12.5 kDa.

In another embodiment of the present invention -P ^a' , -P ^a'' and P ^a''' of formula (a) have a molecular weight of about 15 kDa.

In another embodiment of the present invention -P ^a' , -P ^a'' and P ^a''' of formula (a) have a molecular weight of about 20 kDa.

In one embodiment of the present invention, -Z contains one constituent of formula (a).

In another embodiment of the present invention, -Z contains two constituents of formula (a).

In another embodiment of the present invention, -Z contains three constituents of formula (a).

Preferably, -Z is a constituent of formula (a).

More preferably, -Z contains a term of formula (b)

(b),

(b)

where

the dotted line shows attachment to -L ² - or to the remainder of -Z; and

m and p independently of each other represent an integer in the range and inclusive of 150 to 1000; preferably an integer in the range and inclusive of 150 to 500; more preferably an integer in the range and inclusive of 200 to 500; and most preferably an integer in the range and inclusive of 400 to 500.

Preferably, m and p of formula (b) are the same integer.

Most preferably, m and p of formula (b) are about 450.

Preferably, -Z is a component of formula (b).

The -Z' carrier is a water-insoluble polymer, even more preferably a hydrogel. Preferably, such a hydrogel contains a polymer selected from the group consisting of 2-methacryloyloxyethyl phosphoylcholines, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy)polymers, poly(amides), poly(amidoamines). ), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides) , poly(esters), poly(ethylenes), poly(ethylene glycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazolines), poly(iminocarbonates), poly(lactic acids), poly(lactic-glycolic acid copolymers), poly(methacrylamides), poly(methacrylates) , poly(methyloxazolines), poly(organophosphazenes), poly(orthocompound ethers), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinylamines), poly(vinylmethyl ethers), poly(vinyl pyrrolidones), silicones, celluloses , carbomethylcelluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other polymers based on carbohydrates, xylans and their copolymers .

If the -Z' carrier is a hydrogel, it is preferably a hydrogel containing PEG or hyaluronic acid. Most preferably, such a hydrogel contains PEG.

Even more preferably, the -Z' carrier is a hydrogel as disclosed in WO 2006/003014 A2, WO 2011/012715 A1 or WO 2014/056926 A1, which are incorporated herein by reference in their entirety.

In another embodiment of the present invention, -Z' is a polymer network formed by physical aggregation of polymer chains, the physical aggregation being preferably hydrogen bonded, crystallized, helicalized or complexed. In one embodiment of the present invention, such a polymeric network is a thermogel polymer.

If the PTH controlled release compound for use according to the present invention is a prodrug, its total mass is preferably at least 10 kDa, such as at least 12 kDa, such as at least 15 kDa, such as at least 20 kDa, or as for example at least 30 kDa. If the PTH controlled release compound is a water-soluble prodrug, its total weight is preferably at most 250 kDa, such as at most 200 kDa, 180 kDa, 150 kDa or 100 kDa. It is understood that no significant upper molecular weight limit can be achieved if the controlled release PTH compound is water insoluble.

In one preferred embodiment of the present invention, the PTH controlled release compound has the formula (IIe-i):

(IIe-i),

(IIe-i),

where

the unmarked dotted line denotes the attachment to the nitrogen atom of the -D moiety, which is the PTH moiety, through the formation of an amide bond; and

dashed line marked with an asterisk indicates attachment to component

where

m and p are independently an integer in the range and including 400 - 500.

Preferably, -D is attached to the PTH prodrug of formula (IIe-i) via the N-terminal amine functional group of the PTH moiety.

In another preferred embodiment of the present invention, the PTH prodrug of the present invention has the formula (IIf-i):

(IIf-i),

(IIf-i),

where

the unmarked dotted line denotes the attachment to the nitrogen atom of the -D moiety, which is the PTH moiety, through the formation of an amide bond; and

dashed line marked with an asterisk indicates attachment to component

where

m and p are independently an integer in the range and including 400 - 500.

Preferably, -D is attached to the PTH prodrug of formula (IIf-i) via the N-terminal amine functional group of the PTH moiety.

In a preferred embodiment of the present invention, the residual PTH prodrug activity is less than 10%, more preferably less than 1%, even more preferably less than 0.1%, even more preferably less than 0.01%, even more preferably less than 0.001%, and most preferably less than 0.0001%.

As used herein, the term “residual activity” refers to the activity exhibited by a PTH prodrug wherein the PTH moiety is bound to a carrier relative to the activity exhibited by the corresponding free PTH. In this context, the term "activity" refers to binding to the activation domain of the PTH/PTHrP1 receptor leading to activation of adenylate cyclase to form cAMP, phospholipase C to form intracellular calcium, or osteoblastic expression of RANKL (which binds to RANK (nuclear factor receptor kB activator) on osteoclasts. It is understood that the measurement of residual PTH activity of a prodrug for use according to the present invention requires time during which a certain amount of PTH can be released from the PTH prodrug, and that such released PTH may skew the results measured for the PTH prodrug. test the residual activity of a prodrug with a conjugate in which the drug moiety, in this case PTH, is irreversible, i.e. stably associated with a carrier that, as far as possible, resembles the structure of the PTH prodrug for which residual activity is to be measured.

Preferably, a pharmaceutical composition containing at least one controlled release PTH for use according to the present invention has a pH value in the range and including pH 3 - pH 8. More preferably, the pharmaceutical composition has a pH value in the range and including pH 4 - pH 6. Most preferably, the pharmaceutical composition has a pH value in the range and including pH 4 - pH 5.

In one embodiment of the present invention, the pharmaceutical composition containing at least one controlled release PTH for use according to the present invention is a liquid composition or a composition in the form of a suspension. It is understood that the pharmaceutical composition is a composition in the form of a suspension if the controlled release PTH compound for use according to the present invention is insoluble in water.

In another embodiment of the present invention, the pharmaceutical composition containing at least one controlled release PTH for use according to the present invention is a dry composition that is reconstituted prior to administration to a patient.

Such a liquid pharmaceutical composition, suspension pharmaceutical composition, dry or reconstituted pharmaceutical composition contains at least one excipient. Excipients used in parenteral compositions can be categorized as follows, for example, buffering agents, isotonicity modifiers, preservatives, stabilizers, anti-adsorption agents, anti-oxidants, thickeners/viscosifiers, or other adjuvants. However, in some cases, one excipient may have double or triple functions. Preferably, at least one excipient contained in the pharmaceutical composition for use according to the present invention is selected from the group consisting of the following:

(i) Buffering agents: physiologically acceptable buffers to maintain the pH value in the desired range, such as sodium phosphate, bicarbonate, succinate, histidine, citrate and acetate, sulfate, nitrate, chloride, pyruvate; acid neutralizers such as Mg(OH) ₂ or ZnCO ₃ may also be used;

(ii) Isotonicity modifiers: to minimize pain that may result from cellular damage due to osmotic pressure differences when injected with depot agents; glycerin and sodium chloride are examples; effective concentrations can be determined by osmometry, using an allowable osmotic concentration of the solution equal to 285-315 mOsmol/kg for serum;

(iii) Preservatives and/or antimicrobials: multi-dose parenteral formulations require the addition of preservatives at a sufficient concentration to minimize the risk of infection to the patient upon injection, and appropriate regulatory requirements must be established; exemplary preservatives include m-cresol, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, chlorobutanol, benzyl alcohol, phenylmercury nitrate, thimerosol, sorbic acid, potassium sorbate, benzoic acid, chlorocresol, and benzalkonium chloride;

(iv) Stabilizers: Stabilization is achieved by enhancing protein stabilizing forces, destabilizing the denatured state, or direct binding of excipients to the protein; stabilizers can be amino acids such as alanine, arginine, aspartic acid, glycine, histidine, lysine, proline, sugars such as glucose, sucrose, trehalose, polyols such as glycerol, mannitol, sorbitol, salts such as potassium phosphate, sulfate sodium, chelating agents such as EDTA, hexaphosphate, ligands such as divalent metal ions (zinc, calcium, etc.), other salts or organic molecules such as phenolic derivatives; in addition, oligomers or polymers such as cyclodextrins, dextran, dendrimers, PEG or PVP or protamine or HSA can be used;

(v) Anti-adsorption agents: Generally, ionic or non-ionic surfactants or other proteins or soluble polymers are applied to coat or competitively adsorb to the interior surface of the composition container. For example, poloxamer (Pluronic F-68), PEG dodecyl ether (Brij 35), polysorbate 20 and 80, dextran, polyethylene glycol, PEG-polyhistidine, BSA and HSA, and gelatins; the selected concentration and type of excipient depends on the effect to be avoided, but usually a monolayer of surfactant forms at the interface just above the CMC value;

(vi) Anti-oxidation agents: antioxidants such as ascorbic acid, ectoine, methionine, glutathione, monothioglycerol, morine, polyethyleneimine (PEI), propyl gallate, and vitamin E; chelating agents such as citric acid, EDTA, hexaphosphate and thioglycolic acid may also be used;

(vii) Thickeners or viscosifiers: in the case of a suspension, they slow down the settling of particles in the vial and syringe and are used to facilitate mixing and resuspension of the particles and to facilitate the administration of the suspension (i.e. with little force on the syringe plunger); suitable thickeners or viscosity enhancers are, for example, carbomer thickeners such as Carbopol 940, Carbopol Ultrez 10, cellulose derivatives such as hydroxypropyl methylcellulose (hypromellose, HPMC) or diethylaminoethylcellulose (DEAE or DEAE-C), colloidal magnesium silicate (Veegum) or silicate sodium, hydroxyapatite gel, tricalcium phosphate gel, xanthans, carrageenans such as Satia UTC 30 gum, aliphatic poly(hydroxy acids) such as poly(D, L- or L-lactic acid) (PLA) and poly(glycolic acid) (PGA ) and their copolymers (PLGA), terpolymers of D, L-lactide, glycolide and caprolactone, poloxamers, hydrophilic poly(oxyethylene) blocks and hydrophobic poly(oxypropylene) blocks to create a poly(oxyethylene)-poly(oxypropylene)-poly(oxyethylene) triblock ) (e.g. Pluronic®), a polyether-polyester copolymer such as polyethylene glycol terephthalate/polybutylene terephthalate copolymer, sucrose acetate isobutyrate (SAIB), dextran or its derivatives, combinations of dextrans and PEG, polydimethylsiloxane, collagen, chitosan, polyvinyl alcohol (PVA) and derivatives, polyalkylimides, poly(acrylamide-co-diallyldimethylammonium (DADMA)), polyvinylpyrrolidone (PVP), glucosaminoglycans (GAG) such as dermatan sulfate , chondroitin sulfate, keratan sulfate, heparin, heparan sulfate, hyaluronan, ABA triblock or AB block copolymers consisting of hydrophobic A-blocks, such as polylactide (PLA) or poly(lactide-co-glycolide) (PLGA) and hydrophilic B-blocks, such as polyethylene glycol (PEG) or polyvinylpyrrolidone; such block copolymers, like the aforementioned poloxamers, can exhibit reversible thermal gelation (liquid state at room temperature for ease of administration and gel state above the sol-gel transition temperature at body temperature after injection);

(viii) Lyophilizing or diffusing agent: modifies connective tissue permeability by hydrolyzing extracellular matrix components in the interstitial space such as, but not limited to, hyaluronic acid, a polysaccharide found in the interstitial space of connective tissue; a lyophilizing agent, such as, but not limited to, hyaluronidase, temporarily reduces the viscosity of the extracellular matrix and promotes diffusion of injected drugs; and

(ix) Other adjuvants: such as wetting agents, viscosity modifiers, antibiotics, hyaluronidase; acids and bases such as hydrochloric acid and sodium hydroxide are aids needed to adjust the pH during production.

Another object of the present invention is a method of treating, controlling, slowing down or preventing, in a mammalian patient, one or more conditions that can be treated, controlled, slowed down or prevented by PTH, comprising the step of administering a pharmaceutical composition containing at least one PTH controlled compound. release or a pharmaceutically acceptable salt, hydrate or solvate thereof, not more frequently than once every 24 hours with a dose of a controlled release PTH compound that corresponds to no more than 70% of the molar equivalent dose of PTH 1-84 administered at the same dosing frequency required to maintain serum calcium levels within normal levels for 24 hours.

Preferably, the mammalian patient is a human patient.

Preferred embodiments, for example, controlled release PTH compounds, frequency of administration, i.e. time between two injections, route of administration and dosage, are as described above.

Preferably, the condition that can be treated, controlled, retarded, or prevented by PTH is selected from the group consisting of hypoparathyroidism, hyperphosphatasemia, osteoporosis, fracture healing, bone softening, bone softening, and osteoporosis in patients with hypophosphatasia, steroid-induced osteoporosis, male osteoporosis, arthritis, osteoarthritis, brittle bone syndrome, fibrous dysplasia, rheumatoid arthritis, Paget's disease, malignancy-associated humoral hypercalcemia, osteopenia, periodontal disease, bone fracture, alopecia, chemotherapy-induced alopecia, and thrombocytopenia. More preferably, the condition that can be treated, controlled, retarded, or prevented by PTH is selected from the group consisting of hypoparathyroidism, hyperphosphatasemia, fissure healing, arthritis, osteoarthritis, rheumatoid arthritis, osteopenia, periodontal disease, bone fracture, chemotherapy-induced alopecia alopecia and thrombocytopenia.

Most preferably, said condition is hypoparathyroidism.

Examples

Substances and Methods

Side chain protected PTH(1-34) (SEQ ID NO:51) on TCP resin having Boc protecting group at N-terminus and ivDde protecting Lys26 side chain (synthesized according to Fmoc strategy) obtained from standard peptide synthesis vendors .

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus (synthesized according to the Fmoc strategy) was obtained from standard peptide synthesis vendors.

PEG 2x20 kDa maleimide, Sunbright GL2-400MA was purchased from NOF Europe N.V., Grobbendonk, Belgium. S-Trityl-6-mercaptohexanoic acid was purchased from Polypeptide, Strasbourg, France. HATU was obtained from Merck Biosciences GmbH, Schwalbach/Ts, Germany. Fmoc-N-Me-Asp(OBn)-OH was obtained from Peptide International Inc., Louisville, KY, USA. Fmoc-Aib-OH was purchased from Iris Biotech GmbH, Marktredwitz, Germany. All other chemicals and reagents were purchased from Sigma Aldrich GmbH, Taufkirchen, Germany, unless otherwise specified.

Compound 11a (examples 11-15) was synthesized according to the procedure described in example WO29095479A2, example 1.

Syringes equipped with a polyethylene frit (MultiSynTech GmbH, Witten, Germany) were used as reaction vessels or in the washing step of the peptide resins.

General procedure for deprotecting ivDde of side chain protected PTH on resin: The resin was pre-swollen in DMF for 30 minutes and the solvent was removed. The ivDde group was removed by incubating the resin with DMF/hydrazine hydrate 4/1 (v/v, 2.5 ml/g resin) for 8 x 15 min. A fresh solution of DMF/hydrazine hydrate was used for each step. Finally, the resin was washed with DMF (10x), DCM (10x) and dried in vacuo.

General procedure for deprotection of the Fmoc protecting group from protected PTH on resin: the resin was pre-swollen in DMF for 30 min and the solvent was removed. The Fmoc group was removed by incubating the resin with DMF/piperidine/DBU 96/2/2 (v/v/v, 2.5 ml/g resin) for 3 x 10 min. A fresh solution of DMF/piperidine/DBU was used for each step. Finally, the resin was washed with DMF (10x), DCM (10x) and dried in vacuo.

Purification by RP-HPLC:

Preparative RP-HPLC used a Waters 600 controller and 2487 Dual Absorbance detector equipped with the following columns: Waters XBridge ^TM BEH300 Prep C18 5 µm, 150 x 10 mm, flow rate 6 ml/min, or Waters XBridge ^TM BEH300 Prep C18 10 µm , 150 x 30 mm, flow rate 40 ml/min. Linear gradients of solvent system A (water containing 0.1% TFA v/v or 0.01% conc HCl v/v) and solvent system B (acetonitrile containing 0.1% TFA v/v or 0.01% conc) were applied. .HCl v/v). HPLC fractions containing product were pooled and lyophilized unless otherwise indicated.

Flash chromatography

Flash chromatography purifications were performed on an Isolera One system from Biotage AB, Sweden using Biotage KP-Sil silica gel cartridges and n-heptane and ethyl acetate as eluents. Products were detected at 254 nm.

Ion exchange chromatography:

Ion exchange chromatography (IEX) was performed using an Amersham Bioscience AEKTAbasic AERTAbasic system equipped with a MacroCap SP cation exchange column (Amersham Bioscience/GE Healthcare). The mobile phases used were 17 mM acetic acid, pH 4.5 (solvent A) and 17 mM acetic acid, 1 M NaCl, pH 4.5 (solvent B).

Size exclusion chromatography:

Size exclusion chromatography (SEC) was performed using an Amersham Bioscience AEKTAbasic system equipped with a HiPrep 26/10 desalting column (Amersham Bioscience/GE Healthcare). 0.1% (v/v) acetic acid was used as the mobile phase.

Analytical Methods

Analytical Ultra High Performance LC (UPLC)-MS was performed on a Waters Acquity system equipped with a Waters BEH300 C18 column (particle size 2.1 x 50 mm, particle size 1.7 µm, flow: 0.25 ml/min, solvent A: water, containing 0.04% TFA (v/v), Solvent B: acetonitrile containing 0.05% TFA (v/v) connected to a Thermo Scientific LTQ Orbitrap Discovery mass spectrometer or connected to Waters Microsass ZQ.

Quantitative measurements of serum calcium (sCa), urinary calcium, and serum phosphorus (sP) were performed on a Roche-Hitachi P800 modular chemistry instrument.

Example 1

Synthesis of linker reagent 1f

Linker reagent 1f was synthesized according to the following scheme:

2,4,6- _{trimethoxybenzaldehyde} (2.08 g, 10.61 mmol) was added in parts . The mixture was stirred at room temperature for 90 minutes, acidified with 3 M HCl (4 ml) and stirred for an additional 15 minutes. The reaction mixture was added to saturated NaHCO ₃ (200 ml) and extracted 5x with DCM. The combined organic phases were dried over Na ₂ SO ₄ and the solvents were evaporated in vacuo. The resulting N-methyl-N-Boc-N'-Tmob-ethylenediamine 1a was dried under high vacuum and used in the next step without further step.

Yield: 3.76 g (11.48 mmol, 89% pure, 1a: double Tmob protected product=8:1)

MS: m/z 355.22=[M+H] ⁺ , (calculated monoisotopic mass=354.21).

To a solution of 1a (2 g, 5.65 mmol) in CH ₂ Cl ₂ (24 ml) COMU (4.84 g, 11.3 mmol), N-Fmoc-N-Me-Asp(OBn)-OH (2.08 g, 4.52 mmol) and 2,4,6-collidine (2.65 ml, 20.34 mmol) was added. The reaction mixture was stirred for 3 h at room temperature, diluted with CH ₂ Cl ₂ (250 ml) and washed 3 x with 0.1 MH ₂ SO ₄ (100 ml) and 3 x with brine (100 ml). The aqueous phases were re-extracted with CH ₂ Cl ₂ (100 ml). The combined organic phases were dried over Na ₂ SO ₄ , filtered and the residue was concentrated to a volume of 24 ml. Compound 1b was purified using flash chromatography.

Yield: 5.31 g (148%, 6.66 mmol)

MS: m/z 796.38=[M+H] ⁺ , (calculated monoisotopic mass=795.37).

To a solution of 1b (5.31 g, max. 4.52 mmol, designated N-Fmoc-N-Me-Asp(OBn)-OH) in THF (60 ml) DBU (1.8 ml, 3% v/v) was added. The solution was stirred for 12 min at room temperature, diluted with DCM (400 ml) and washed 3x with 0.1 M H₂SO_four (150 ml) and 3x with saline (150 ml). The aqueous phases were re-extracted with DCM (100 ml). The combined organic phases were dried over Na₂SO_four and filtered out. Compound 1c was isolated by evaporation of the solvent and used in the next step without further purification.

MS: m/z 574.31=[M+H] ⁺ , (calculated monoisotopic mass=573.30).

Compound 1c (5.31 g, 4.52 mmol, crude) was dissolved in acetonitrile (26 mL) and COMU (3.87 g, 9.04 mmol), 6-tritylmercaptocaproic acid (2.12 g, 5.42 mmol) and 2,4,6-collidine (2.35 mL , 18.08 mmol) was added. The reaction mixture was stirred for 4 h at room temperature, diluted with DCM (400 ml) and washed 3x with 0.1 M H₂SO_four (100 ml) and 3x with saline (100 ml). The aqueous phases were re-extracted with DCM (100 ml). The combined organic phases were dried over Na₂SO_four, filtered and 1d was isolated by evaporating the solvent. Product 1d was purified using flash chromatography.

Yield: 2.63 g (62%, 94% pure)

MS: m/z 856.41=[M+H] ⁺ , (calculated monoisotope mass=855.41).

To a solution of 1d (2.63 g, 2.78 mmol) in i-PrOH (33 ml) and H₂O (11 ml) was added LiOH (267 mg, 11.12 mmol) and the reaction mixture was stirred for 70 min at room temperature. The mixture was diluted with DCM (200 ml) and washed 3x with 0.1 M H₂SO_four (50 ml) and 3x with saline (50 ml). The aqueous phases were re-extracted with DCM (100 ml). The combined organic phases were dried over Na₂SO_four, filtered, and compound 1e was isolated by evaporating the solvent. Compound 1e was purified using flash chromatography.

Yield: 2.1 g (88%)

MS: m/z 878.4=[M+Na] ⁺ , (calculated monoisotope mass=837.40).

To a solution of 1e (170 mg, 0.198 mmol) in anhydrous DCM (4 mL) was added DCC (123 mg, 0.59 mmol) and a catalytic amount of DMAP. After 5 min N-hydroxy-succinimide (114 mg, 0.99 mmol) was added and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was filtered, the solvent was removed in vacuo and the residue was dissolved in 90% acetonitrile plus 0.1% TFA ( 3.4 ml). The crude mixture was purified by RP-HPLC. Product fractions were neutralized with 0.5 M pH 7.4 phosphate buffer and concentrated. The remaining aqueous phase was extracted with DCM and 1f was isolated by evaporating the solvent.

Yield: 154 mg (81%)

MS: m/z 953.4=[M+H] ⁺ , (calculated monoisotope mass=952.43).

Example 2

Synthesis of linker reagent 2g

4-Methoxytriphenylmethyl chloride (3.00 g, 9.71 mmol) was dissolved in DCM (20 ml) and added dropwise with stirring to a solution of ethylenediamine 2a (6.5 ml, 97.3 mmol) in DCM (20 ml). The reaction mixture was stirred for 2 h at room temperature, after which it was diluted with diethyl ether (300 ml), washed 3x with brine/0.1 M NaOH 30/1 (v/v) and once with brine. The organic phase was dried over Na ₂ SO ₄ and compound 2b was isolated by evaporating the solvent.

Yield: 3.18 g (98%)

Mmt-protected intermediate 2b (3.18 g, 9.56 mmol) was dissolved in DCM (30 ml). 6-(Tritylthio)-hexanoic acid (4.48 g, 11.5 mmol), PyBOP (5.67 g, 10.9 mmol) and DIPEA (5.0 ml, 28.6 mmol) were added and the mixture was stirred for 30 minutes at room temperature. The solution was diluted with diethyl ether (250 ml), washed 3x with brine/0.1 M NaOH 30/1 (v/v) and once with brine. The organic phase was dried over Na ₂ SO ₄ and the solvent was removed in vacuo. 2c was purified using flash chromatography.

Yield: 5.69 g (85%)

MS: m/z 705.4=[M+H] ⁺ , (calculated monoisotope mass=704.34).

Compound 2c (3.19 g, 4.53 mmol) was dissolved in anhydrous THF (50 mL), a 1 M BH ₃ ·THF solution in THF (8.5 mL, 8.5 mmol) was added and the mixture was stirred for 16 h at room temperature. Another 1 M BH ₃ ·THF solution in THF (14 ml, 14.0 mmol) was added and the mixture was stirred for another 16 h at room temperature. Methanol (8.5 ml) and N,N'-dimethyl-ethylenediamine (3.00 ml, 27.9 mmol) were added and the mixture was heated under reflux for 3 hours. The mixture was allowed to cool and ethyl acetate (300 ml) was added. The solution was washed with 2 x aq. Na ₂ CO ₃ and 2 x aq. NaHCO ₃ . The organic phase was dried over Na ₂ SO ₄ and the solvent removed in vacuo to give compound 2d.

Yield: 3.22 g (103%)

MS: m/z 691.4=[M+H] ⁺ , (calculated monoisotopic mass=690.36).

Di-tert-butyl dicarbonate (2.32 g, 10.6 mmol) and DIPEA (3.09 ml, 17.7 mmol) were dissolved in DCM (5 ml) and added to a solution of 2d (2.45 g, 3.55 mmol) in DCM (5 ml). The mixture was stirred for 30 minutes at room temperature. The solution was concentrated in vacuo and purified using flash chromatography to give product 2e.

Yield: 2.09 g (74%)

MS: m/z 791.4=[M+H] ⁺ , (calculated monoisotopic mass=790.42).

Compound 2e (5.01 g, 6.34 mmol) was dissolved in acetonitrile (80 ml). 0.4 M aqueous HCl (80 ml) followed by acetonitrile (20 ml) was added and the mixture was stirred for 1 h at room temperature. The pH was adjusted to pH 5.5 by adding aqueous 5 M NaOH. The organic solvent was removed in vacuo and the remaining aqueous solution was extracted 4x with DCM. The combined organic phases were dried over Na ₂ SO ₄ and the solvent removed in vacuo to give product 2f.

Yield: 4.77 g (95%)

MS: m/z 519.3=[M+H] ⁺ , (calculated monoisotopic mass=518.30).

Compound 2f (5.27 g, 6.65 mmol) was dissolved in DCM (30 ml) and added to a solution of p-nitrophenyl fluoroformate (2.01 g, 9.98 mmol) in DCM (25 ml). 2,4,6-trimethylpyridine (4.38 ml, 33.3 mmol) was added and the solution was stirred for 45 minutes at room temperature. The solution was concentrated in vacuo and purified using flash chromatography to give the product 2g.

Yield: 4.04 g (89%)

MS: m/z 706.32=[M+Na] ⁺ , (calculated monoisotope mass=683.30).

Example 3

Synthesis of permanent conjugate S1 PTH(1-34) 3

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus was subjected to Fmoc deprotection according to the procedure described in the Substances and Methods part. A solution of 6-tritylmercaptohexanoic acid (62.5 mg, 160 µmol), PyBOP (80.1 mg, 154 µmol) and DIPEA (53 µl, 306 µmol) in DMF (2 ml) was added to 0.21 g (51 µmol) of the resin. The suspension was shaken for 80 minutes at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 10 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 3 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 36 mg (14%), 3*8 TFA

MS: m/z 1062.31=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1062.30).

Example 4

Synthesis of permanent conjugate K26 PTH(1-34) 4

Side chain protected PTH(1-34) on a TCP resin having a Boc protecting group at the N-terminus and an ivDde protected Lys26 side chain was subjected to ivDde deprotection according to the procedure given in Substances and Methods. A solution of 6-tritylmercaptohexanoic acid (107 mg, 273 µmol), PyBOP (141 mg, 273 µmol) and DIPEA (95 µl, 545 µmol) in DMF (3 ml) was added to 0.80 g (90.9 µmol) of the resin. The suspension was shaken for 1 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 6 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 4 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 40mg (8%), 4*8 TFA

MS: m/z 1062.30=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1062.30).

Example 5

Synthesis of temporary conjugate S1 PTH(1-34)

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus was subjected to Fmoc deprotection according to the procedure described in the Substances and Methods part. A solution of Fmoc-Aib-OH (79 mg, 244 µmol), PyBOP (127 mg, 244 µmol) and DIPEA (64 µl, 365 µmol) in DMF (1.5 ml) was added to 0.60 g (61 µmol) of resin. The suspension was shaken for 16 hours at room temperature. The resin was washed 10x with DMF and subjected to Fmoc deprotection as described above. A solution of compound 2g (167 mg, 244 µmol) and DIPEA (64 µl, 365 µmol) in DMF (1.5 ml) was added to the resin. The suspension was shaken for 24 hours at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 7 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 5 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 78 mg (24%), 5*9 TFA

MS: m/z 1101.59=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1101.57).

Example 6

Synthesis of temporary conjugate S1 PTH(1-34) 6

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus was subjected to Fmoc deprotection according to the procedure described in the Substances and Methods part. A solution of Fmoc-Ala-OH (32 mg, 102 µmol), PyBOP (53 mg, 102 µmol) and DIPEA (27 µl, 152 µmol) in DMF (3 ml) was added to 0.25 g (25 µmol) of resin. The suspension was shaken for 1 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Deprotection of the Fmoc group was carried out as described above. A solution of compound 2g (69 mg, 102 µmol) and DIPEA (27 µl, 152 µmol) in DMF (3 ml) was added to the resin. The suspension was shaken for 1.5 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 3 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 6 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 25 mg (18%), 6*9 TFA

MS: m/z 1098.75=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1098.07).

Example 7

Synthesis of temporary conjugate S1 PTH(1-34) 7

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus was subjected to Fmoc deprotection according to the procedure described in the Substances and Methods part. A solution of Fmoc-Ser(Trt)-OH (117 mg, 205 µmol), PyBOP (108 mg, 207 µmol) and DIPEA (53 µl, 305 µmol) in DMF (2 ml) was added to 0.50 g (51 µmol) of the resin. The suspension was shaken for 1 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Deprotection of the Fmoc group was carried out as described above. A solution of 2g (144 mg, 211 µmol) and DIPEA (53 µl, 305 µmol) in DMF (1.8 ml) was added to the resin. The suspension was shaken for 7 hours at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection of the protecting groups was achieved by adding 6 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 7 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 54 mg (20%), 7*9 TFA

MS: m/z 1102.08=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1102.07).

Example 8

Synthesis of temporary conjugate S1 PTH(1-34) 8

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus was subjected to Fmoc deprotection according to the procedure described in the Substances and Methods part. A solution of Fmoc-Leu-OH (36 mg, 102 µmol), PyBOP (53 mg, 102 µmol) and DIPEA (27 µl, 152 µmol) in DMF (3 ml) was added to 0.25 g (25 µmol) of resin. The suspension was shaken for 1 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Deprotection of the Fmoc group was carried out as described above. A solution of compound 2g (69 mg, 102 µmol) and DIPEA (27 µl, 152 µmol) in DMF (3 ml) was added to the resin. The suspension was shaken for 1.5 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 3 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 8 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 31 mg (22%), 8*9 TFA

MS: m/z 1109.32=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1108.58).

Example 9

Synthesis of temporary conjugate S1 PTH(1-34) 9

Side chain protected PTH(1-34) on a TCP resin having an Fmoc protecting group at the N-terminus was subjected to Fmoc deprotection according to the procedure described in the Substances and Methods part. A solution of compound 1e (182 mg, 213 µmol), PyBOP (111 mg, 213 µmol) and DIPEA (93 µl, 532 µmol) in DMF (5 ml) was added to 2.00 g (107 µmol) of resin. The suspension was shaken for 16 hours at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 20 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 9 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 47 mg (8%), 9*9 TFA

MS: m/z 1108.58=[M+4H] ⁴⁺ , (calculated monoisotopic mass for [M+4H] ⁴⁺ =1108.57).

Example 10

Synthesis of temporary conjugate K26 PTH(1-34) 10

Side chain protected PTH(1-34) on a TCP resin having a Boc protecting group at the N-terminus and an ivDde protected Lys26 side chain was subjected to ivDde deprotection according to the procedure given in Substances and Methods. A solution of compound 1f (867 mg, 910 µmol) and DIPEA (0.24 ml, 1.36 mmol) in DMF (5 ml) was added to 1.91 g (227 µmol) of resin. The suspension was shaken for 1 h at room temperature. The resin was washed 10x with DMF, 10x with DCM and dried in vacuo. Cleavage of the peptide from the resin and deprotection was achieved by adding 20 ml of cleavage cocktail 100/3/3/2/1 (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for 1 h at room temperature. The crude compound 10 was precipitated in pre-chilled simple diethyl ether (-18°C). The precipitate was dissolved in ACN/water and purified by RP-HPLC. Product fractions were lyophilized.

Yield: 92 mg (7%), 10*9 TFA

MS: m/z 1108.58=[M+4H] ⁴⁺ , (calculated monoisotopic mass for [M+4H] ⁴⁺ =1108.57).

Example 11

Synthesis of low molecular weight temporary conjugate S1 PEG 11b

0.15 ml of 0.5 M NaH ₂ PO ₄ buffer (pH 7.4) was added to 0.5 ml of a 20 mg/ml solution of thiol 5 (10 mg, 1.84 µmol) in 1/1 (v/v) acetonitrile/water containing 0.1% TFA (vol./vol.). The solution was incubated at room temperature for 10 minutes, after which 238 µl of a 10 mg/ml solution of maleimide 11a (2.4 mg, 2.21 µmol) in 1/1 (v/v) acetonitrile/water containing 0.1% TFA (v/v) /rev.), added. The solution was incubated for 20 minutes at room temperature. 10 μl TFA was added and the mixture was purified by RP-HPLC. Product fractions were lyophilized to give compound 11b.

Yield: 3.1 mg (26%), 11b*9 TFA

MS: m/z 1097.00=[M+4H] ⁴⁺ , (calculated monoisotopic mass for [M+5H] ⁵⁺ =1096.99).

Example 12

Synthesis of low molecular weight temporary conjugate S1 PEG 12

Conjugate 12 was synthesized as described for 11b using thiol 6 (10 mg, 1.85 µmol) and maleimide 11a (2.4 mg, 2.21 µmol).

Yield: 10 mg (83%), 12*9 TFA

MS: m/z 1094.20=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1094.19).

Example 13

Synthesis of low molecular weight temporary conjugate S1 PEG 13

Conjugate 13 was synthesized as described for 11b using thiol 7 (10 mg, 1.84 µmol) and maleimide 11a (2.4 mg, 2.21 µmol).

Yield: 8 mg (67%), 13*9 TFA

MS: m/z 1097.40=[M+5H] ⁵⁺ , (calculated monoisotope mass for [M+5H] ⁵⁺ =1097.39).

Example 14

Synthesis of low molecular weight temporary conjugate S1 PEG 14

Conjugate 14 was synthesized as described for 11b using thiol 8 (10 mg, 1.83 µmol) and maleimide 11a (2.4 mg, 2.21 µmol).

Yield: 4 mg (33%), 14*9 TFA

MS: m/z 1378.01=[M+4H] ⁴⁺ , (calculated monoisotope mass for [M+4H] ⁴⁺ =1378.00).

Example 15

Synthesis of low molecular weight temporary conjugate K26 PEG 15

Conjugate 15 was synthesized as described for 11b using thiol 10 (5.2 mg, 0.95 µmol) and maleimide 11a (1.23 mg, 1.14 µmol).

Yield: 2.1 mg (33%), 15*9 TFA

MS: m/z 1102.60=[M+5H] ⁵⁺ , (calculated monoisotope mass for [M+5H] ⁵⁺ =1102.59).

Example 16

Synthesis of permanent conjugate 2x20 kDa S1 PEG 16

772 µl of a solution containing thiol 3 (19.4 mg/ml, 15 mg, 3.54 µmol) and 2.5 mg/ml Boc-L-Met in 1/1 (v/v) acetonitrile/water containing 0.1% TFA (v/v) ./vol.), was added to 1.87 ml of a solution containing PEG 2x20 kDa - maleimide (Sunbright GL2-400MA, 187 mg, 4.32 µmol) and 2.5 mg/ml Boc-L-Met in water containing 0.1% TFA (vol. /about.). 0.5 M NaH ₂ PO ₄ buffer (0.66 ml, pH 7.0) was added and the mixture was stirred for 30 minutes at room temperature. 10 μl of a 270 mg/ml solution of 2-mercaptoethanol in water was added. The mixture was stirred for 5 minutes at room temperature and 0.33 ml of 1 M HCl was added. Conjugate 16 was purified by IEX followed by RP-HPLC using a linear gradient of solvent system A (water containing 0.1% AcOH v/v) and solvent system B (acetonitrile containing 0.1% AcOH v/v). Fractions containing the solution were lyophilized.

Yield: 97 mg (2.01 µmol, 57%) of 16*8 AcOH conjugate

Example 17

Synthesis of permanent conjugate 2x20 kDa K26 PEG 17

Conjugate 17 was prepared as described for 16 by reacting thiol 4 (15 mg, 3.53 µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, 187 mg, 4.32 µmol).

Yield: 80 mg (1.79 µmol, 51%) of 17*8 AcOH conjugate

Example 18

Synthesis of temporary conjugate 2x20 kDa S1 PEG 18

Conjugate 18 was prepared as described for 16 by reacting thiol 5 (37 mg, 8.40 µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, 445 mg, 9.24 µmol). The reaction was quenched by the addition of 50 μl of TFA without the addition of 2-mercaptoethanol. Conjugate 18 was purified by IEX followed by SEC for desalting. Fractions containing the solution were lyophilized.

Yield: 161 mg (3.33 µmol, 40%) conjugate 18*9 AcOH

Example 19

Synthesis of temporary conjugate 2x20 kDa S1 PEG 19

Conjugate 19 was prepared as described for 16 by reacting thiol 7 (27 mg, 6.14 µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, 325 mg, 7.50 µmol).

Yield: 249 mg (5.16 µmol, 84%) conjugate 19*9 AcOH

Example 20

Synthesis of temporary conjugate 2x20 kDa S1 PEG 20

Conjugate 20 was prepared as described for 16 by reacting thiol 9 (38 mg, 8.59 µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, 455 mg, 9.45 µmol). The reaction was quenched by the addition of 50 μl of TFA without the addition of 2-mercaptoethanol. Conjugate 20 was purified by IEX followed by SEC for desalting. Fractions containing the solution were lyophilized.

Yield: 194 mg (4.01 µmol, 47%) conjugate 20*9 AcOH

Example 21

Synthesis of temporary conjugate 2x20 kDa K26 PEG 21

Conjugate 21 was prepared as described for 16 by reacting thiol 10 (34 mg, 7.58 µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, 401 mg, 9.26 µmol).

Yield: 256 mg (5.30 µmol, 70%) conjugate 21*9 AcOH

Example 22

Pharmacodynamic actions in thyroparathyroidectomized (TPTx) rats during a 28-day study with daily subcutaneous injections of conjugate 18 or PTH(1-84)

This study was conducted in order to test and compare the effect of daily subcutaneous injection of Compound 18 and PTH (1-84), the current safety standard, in an animal model of the disease relevant to the treatment of hypoparathyroidism (HP) study. Rats that have undergone thyroparathyroidectomy (TPTx) by blunt dissection cannot produce the parathyroid hormone, PTH, a major regulator of calcium homeostasis. Consequently, TPTx rats develop hypocalcemia and hyperphosphatasemia characteristic of HP. 17-week-old female SD TPTx rats (n=9/group) were dosed subcutaneously for 28 days with compound 18 (5 μg PTH eq/kg/day, 1.2 nmol/kg/day, in 10 mM succinic acid, 46 g /l mannitol, pH 4.0), PTH (1-84) (70 μg PTH eq/kg/day, 7.3 nmol/kg/day, in 10 mM citrate, mannitol 39.0 g/l, pH 5 ,0) or environment. Additionally, one group of sham-operated rats (n=9) representing a normal physiological background control also received the medium. Serum calcium (sCa) and phosphorus (sP) levels in animals were measured before and after dosing on days 1, 6, 12 and 27. In addition, markers of bone remodeling (P1NP and CTx) were measured and bone quality was assessed by ex vivo pQCT.

Results: Mean sCa in TPTx rats pre-dosed on day 1 was 8.3 mg/dL compared to 10.9 mg/dL in sham-operated control rats. sP values were 8.7 mg/dL and 5.9 mg/dL. dl, respectively. Compound 18 was administered daily at 1.2 nmol/kg, increasing sCa to nearly normal levels while decreasing sP over several days of administration. On day 12 (day 5 at steady state with compound 18), sCa had stabilized at normal levels (10.7 mg/dl) in this group of animals (compound 18/sham control ratio = 1.01) in contrast to the hypocalcemic level (8, 1 mg/dl) measured in PTH(1-84) treated rats (ratio PTH(1-84)/sham control=0.76). In addition, the 24-hour urinary Ca excretion at day 12 was comparable between Compound 18-treated animals and sham-operated controls. Bone mineral density (BMD) and bone mineral content (BMC) were increased in TPTx controls, as observed in HP patients. Compound 18 treatment reduced BMD, BMC, and area in parallel with an increase in CTx compared to sham-operated and TPTx medium-treated animals. A significant increase in trabecular BMD was observed in PTH(1-84) treated animals compared to both control groups.

It was concluded that Compound 18, at a dose even less than 20% of the molar equivalent of the PTH(1-84) dose tested here, is able to maintain sCa at a level comparable to that of sham-operated control animals (represents the normal level here) in 24 hours. In contrast, PTH(1-84) at 7.3 nmol/kg/day did not result in an increase in sCa compared to the levels of medium-injected TPTx rats. However, a minimal decrease in sP was observed in animals dosed with PTH(1-84), confirming exposure to and response to PTH(1-84) in rats. After 28 days of compound 18 treatment, trabecular and cortical BMD in the vertebrae were normal range, while an anabolic effect was observed for PTH(1-84) on trabecular and cortical bones in the vertebrae.

Abbreviations:

ACN acetonitrile

AcOH acetic acid

Aib 2-aminoisobutyric acid

BMD bone mineral density

Bn benzyl

Boc tert-butyloxycarbonyl

COMU (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate

cAMP cyclic adenosine monophosphate

d day

DBU 1,3-diazabicyclo[5.4.0]undecene

DCC N,N'-dicyclohexylcarbodiimide

DCM dichloromethane

DIPEA N,N-diisopropylethylamine

DMAP dimethylamino-pyridine

DMF N,N-dimethylformamide

DMSO dimethyl sulfoxide

DTT dithiothreitol

EDTA ethylenediaminetetraacetic acid

eq stoichiometric equivalent

ESI-MS Electrospray Ionization Mass Spectrometry

Et ethyl

Fmoc 9-fluorenylmethyloxycarbonyl

Glu-C endoproteinase Glu-C

h hour

HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate

HP hypoparathyroidism

HPLC high performance liquid chromatography

ivDde 4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl

LC liquid chromatography

LTQ linear quadrupole trap

Lys-C endoproteinase Lys-C

LLOQ lower limit of quantity

Mal 3-maleimidopropyl

Me methyl

MeOH methanol

min minutes

Mmt Monomethoxytrityl

MS mass spectrometer / mass spectrometry

m/z mass-charge ratio

OtBu tert-butyloxy

PEG poly(ethylene glycol)

pH potentia Hydrogenii

PK pharmacokinetics

Pr propyl

PTH parathyroid hormone

PyBOP benzotriazol-1-yl-hydroxytripyrrolidinophosphonium hexafluorophosphate

Q-TOF quadrupole TOF

RP-HPLC Reversed Phase High Performance Liquid Chromatography

rt room temperature

sCa serum calcium

SIM monitoring of selected ions

SEC size exclusion chromatography

sc subcutaneously

sP whey phosphate

t _{1/2 half} -life

TCP trityl chloride polystyrene

TES triethylsilane

TFA trifluoroacetic acid

THF tetrahydrofuran

Tmob 2,4,6-trimethoxybenzyl

TPTx thyroparathyroidectomy

Trt triphenylmethyl, trityl

ULOQ Quantitative Upper Limit

UPLC ultra high performance liquid chromatography

UV ultraviolet

ZQ single quadrupole

--->

Sequence listing

<110>Ascendis Pharma A/S

<120>CONTROLLED RELEASE PTH COMPOUND DOSING MODE

<130>CPX70582PC

<160>122

<170>PatentIn version 3.5

<210>1

<211>84

<212>PRT

<213>Homo sapiens

<400>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

<210>2

<211>83

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-83

<400>2

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

<210>3

<211>82

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-82

<400>3

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

<210>4

<211>81

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-81

<400>4

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

<210>5

<211>80

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-80

<400>5

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

<210>6

<211>79

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-79

<400>6

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

65 70 75

<210>7

<211>78

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-78

<400>7

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

65 70 75

<210>8

<211>77

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-77

<400>8

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

65 70 75

<210>9

<211>76

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-76

<400>9

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

65 70 75

<210>10

<211>75

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-75

<400>10

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

65 70 75

<210>11

<211>74

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-74

<400>11

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

65 70

<210>12

<211>73

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-73

<400>12

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

65 70

<210>13

<211>72

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-72

<400>13

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

65 70

<210>14

<211>71

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-71

<400>14

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

65 70

<210>15

<211>70

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-70

<400>15

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

65 70

<210>16

<211>69

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-69

<400>16

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

65

<210>17

<211>68

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-68

<400>17

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

65

<210>18

<211>67

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-67

<400>18

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

65

<210>19

<211>66

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-66

<400>19

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

65

<210>20

<211>65

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-65

<400>20

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

65

<210>21

<211>64

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-64

<400>21

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

<210>22

<211>63

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-63

<400>22

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

50 55 60

<210>23

<211>62

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-62

<400>23

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

50 55 60

<210>24

<211>61

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-61

<400>24

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

50 55 60

<210>25

<211>60

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-60

<400>25

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

50 55 60

<210>26

<211>59

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-59

<400>26

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

50 55

<210>27

<211>58

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-58

<400>27

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

50 55

<210>28

<211>57

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-57

<400>28

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

50 55

<210>29

<211>56

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-56

<400>29

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

50 55

<210>30

<211>55

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-55

<400>30

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

50 55

<210>31

<211>54

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-54

<400>31

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

fifty

<210>32

<211>53

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-53

<400>32

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

fifty

<210>33

<211>52

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-52

<400>33

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

fifty

<210>34

<211>51

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-51

<400>34

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

fifty

<210>35

<211>50

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-50

<400>35

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

fifty

<210>36

<211>49

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-49

<400>36

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

<210>37

<211>48

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-48

<400>37

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

<210>38

<211>47

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-47

<400>38

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

35 40 45

<210>39

<211>46

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-46

<400>39

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

35 40 45

<210>40

<211>45

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-45

<400>40

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

35 40 45

<210>41

<211>44

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-44

<400>41

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

35 40

<210>42

<211>43

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-43

<400>42

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

35 40

<210>43

<211>42

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-42

<400>43

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

35 40

<210>44

<211>41

<212>PRT

<213>artificial sequence

<220>

<223>human PTH-41

<400>44

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

35 40

<210>45

<211>40

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-40

<400>45

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

35 40

<210>46

<211>39

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-39

<400>46

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

35

<210>47

<211>38

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-38

<400>47

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

35

<210>48

<211>37

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-37

<400>48

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

35

<210>49

<211>36

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-36

<400>49

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

35

<210>50

<211>35

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-35

<400>50

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

35

<210>51

<211>34

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-34

<400>51

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

<210>52

<211>33

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-33

<400>52

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

<210>53

<211>32

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-32

<400>53

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

<210>54

<211>31

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-31

<400>54

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

20 25 30

<210>55

<211>30

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-30

<400>55

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

20 25 30

<210>56

<211>29

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-29

<400>56

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

20 25

<210>57

<211>28

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-28

<400>57

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

20 25

<210>58

<211>27

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-27

<400>58

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

20 25

<210>59

<211>26

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-26

<400>59

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

20 25

<210>60

<211>25

<212>PRT

<213>artificial sequence

<220>

<223>human PTH 1-25

<400>60

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

20 25

<210>61

<211>84

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-84

<220>

<221>MOD_RES

<222>(84)..(84)

<223>Amidation

<400>61

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

<210>62

<211>83

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-83

<220>

<221>MOD_RES

<222>(83)..(83)

<223>Amidation

<400>62

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

<210>63

<211>82

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-82

<220>

<221>MOD_RES

<222>(82)..(82)

<223>Amidation

<400>63

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

<210>64

<211>81

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-81

<220>

<221>MOD_RES

<222>(81)..(81)

<223>Amidation

<400>64

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

<210>65

<211>80

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-80

<220>

<221>MOD_RES

<222>(80)..(80)

<223>Amidation

<400>65

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

<210>66

<211>79

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-79

<220>

<221>MOD_RES

<222>(79)..(79)

<223>Amidation

<400>66

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

65 70 75

<210>67

<211>78

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-78

<220>

<221>MOD_RES

<222>(78)..(78)

<223>Amidation

<400>67

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

65 70 75

<210>68

<211>77

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-77

<220>

<221>MOD_RES

<222>(77)..(77)

<223>Amidation

<400>68

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

65 70 75

<210>69

<211>76

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-76

<220>

<221>MOD_RES

<222>(76)..(76)

<223>Amidation

<400>69

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

65 70 75

<210>70

<211>75

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-75

<220>

<221>MOD_RES

<222>(75)..(75)

<223>Amidation

<400>70

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

65 70 75

<210>71

<211>74

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-74

<220>

<221>MOD_RES

<222>(74)..(74)

<223>Amidation

<400>71

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

65 70

<210>72

<211>73

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-73

<220>

<221>MOD_RES

<222>(73)..(73)

<223>Amidation

<400>72

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

65 70

<210>73

<211>72

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-72

<220>

<221>MOD_RES

<222>(72)..(72)

<223>Amidation

<400>73

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

65 70

<210>74

<211>71

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-71

<220>

<221>MOD_RES

<222>(71)..(71)

<223>Amidation

<400>74

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

65 70

<210>75

<211>70

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-70

<220>

<221>MOD_RES

<222>(70)..(70)

<223>Amidation

<400>75

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

65 70

<210>76

<211>69

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-69

<220>

<221>MOD_RES

<222>(69)..(69)

<223>Amidation

<400>76

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

65

<210>77

<211>68

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-68

<220>

<221>MOD_RES

<222>(68)..(68)

<223>amidation

<400>77

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

65

<210>78

<211>67

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-67

<220>

<221>MOD_RES

<222>(67)..(67)

<223>Amidation

<400>78

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

65

<210>79

<211>66

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-66

<220>

<221>MOD_RES

<222>(66)..(66)

<223>Amidation

<400>79

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

65

<210>80

<211>65

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-65

<220>

<221>MOD_RES

<222>(65)..(65)

<223>Amidation

<400>80

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

65

<210>81

<211>64

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-64

<220>

<221>MOD_RES

<222>(64)..(64)

<223>Amidation

<400>81

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

<210>82

<211>63

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-63

<220>

<221>MOD_RES

<222>(63)..(63)

<223>Amidation

<400>82

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

50 55 60

<210>83

<211>62

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-62

<220>

<221>MOD_RES

<222>(62)..(62)

<223>Amidation

<400>83

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

50 55 60

<210>84

<211>61

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-61

<220>

<221>MOD_RES

<222>(61)..(61)

<223>Amidation

<400>84

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

50 55 60

<210>85

<211>60

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-60

<220>

<221>MOD_RES

<222>(60)..(60)

<223>acetylation

<400>85

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

50 55 60

<210>86

<211>59

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-59

<220>

<221>MOD_RES

<222>(59)..(59)

<223>Amidation

<400>86

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

50 55

<210>87

<211>58

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-58

<220>

<221>MOD_RES

<222>(58)..(58)

<223>Amidation

<400>87

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

50 55

<210>88

<211>57

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-57

<220>

<221>MOD_RES

<222>(57)..(57)

<223>Amidation

<400>88

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

50 55

<210>89

<211>56

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-56

<220>

<221>MOD_RES

<222>(56)..(56)

<223>Amidation

<400>89

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

50 55

<210>90

<211>55

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-55

<220>

<221>MOD_RES

<222>(55)..(55)

<223>Amidation

<400>90

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

50 55

<210>91

<211>54

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-54

<220>

<221>MOD_RES

<222>(54)..(54)

<223>Amidation

<400>91

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

fifty

<210>92

<211>53

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-53

<220>

<221>MOD_RES

<222>(53)..(53)

<223>Amidation

<400>92

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

fifty

<210>93

<211>52

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-52

<220>

<221>MOD_RES

<222>(52)..(52)

<223>Amidation

<400>93

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

fifty

<210>94

<211>51

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-51

<220>

<221>MOD_RES

<222>(51)..(51)

<223>Amidation

<400>94

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

fifty

<210>95

<211>50

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-50

<220>

<221>MOD_RES

<222>(50)..(50)

<223>Amidation

<400>95

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

fifty

<210>96

<211>49

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-49

<220>

<221>MOD_RES

<222>(49)..(49)

<223>Amidation

<400>96

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

<210>97

<211>48

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-48

<220>

<221>MOD_RES

<222>(48)..(48)

<223>Amidation

<400>97

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

<210>98

<211>47

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-47

<220>

<221>MOD_RES

<222>(47)..(47)

<223>amidation

<400>98

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

35 40 45

<210>99

<211>46

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-46

<220>

<221>MOD_RES

<222>(46)..(46)

<223>Amidation

<400>99

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

35 40 45

<210>100

<211>45

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-45

<220>

<221>MOD_RES

<222>(45)..(45)

<223>Amidation

<400>100

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

35 40 45

<210>101

<211>44

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-44

<220>

<221>MOD_RES

<222>(44)..(44)

<223>Amidation

<400>101

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

35 40

<210>102

<211>43

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-43

<220>

<221>MOD_RES

<222>(43)..(43)

<223>Amidation

<400>102

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

35 40

<210>103

<211>42

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-42

<220>

<221>MOD_RES

<222>(42)..(42)

<223>Amidation

<400>103

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

35 40

<210>104

<211>41

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-41

<220>

<221>MOD_RES

<222>(41)..(41)

<223>Amidation

<400>104

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

35 40

<210>105

<211>40

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-40

<220>

<221>MOD_RES

<222>(40)..(40)

<223>Amidation

<400>105

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

35 40

<210>106

<211>39

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-39

<220>

<221>MOD_RES

<222>(39)..(39)

<223>Amidation

<400>106

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

35

<210>107

<211>38

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-38

<220>

<221>MOD_RES

<222>(38)..(38)

<223>Amidation

<400>107

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

35

<210>108

<211>37

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-37

<220>

<221>MOD_RES

<222>(37)..(37)

<223>Amidation

<400>108

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

35

<210>109

<211>36

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-36

<220>

<221>MOD_RES

<222>(36)..(36)

<223>Amidation

<400>109

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

35

<210>110

<211>35

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-35

<220>

<221>MOD_RES

<222>(35)..(35)

<223>Amidation

<400>110

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

35

<210>111

<211>34

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-34

<220>

<221>MOD_RES

<222>(34)..(34)

<223>Amidation

<400>111

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

<210>112

<211>33

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-33

<220>

<221>MOD_RES

<222>(33)..(33)

<223>Amidation

<400>112

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

<210>113

<211>32

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-32

<220>

<221>MOD_RES

<222>(32)..(32)

<223>Amidation

<400>113

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

<210>114

<211>31

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-31

<220>

<221>MOD_RES

<222>(31)..(31)

<223>Amidation

<400>114

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

20 25 30

<210>115

<211>30

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-30

<220>

<221>MOD_RES

<222>(30)..(30)

<223>Amidation

<400>115

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

20 25 30

<210>116

<211>29

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-29

<220>

<221>MOD_RES

<222>(29)..(29)

<223>Amidation

<400>116

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

20 25

<210>117

<211>28

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-28

<220>

<221>MOD_RES

<222>(28)..(28)

<223>Amidation

<400>117

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

20 25

<210>118

<211>27

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-27

<220>

<221>MOD_RES

<222>(27)..(27)

<223>Amidation

<400>118

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

20 25

<210>119

<211>26

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-26

<220>

<221>MOD_RES

<222>(26)..(26)

<223>Amidation

<400>119

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

20 25

<210>120

<211>25

<212>PRT

<213>artificial sequence

<220>

<223>amidated human PTH 1-25

<220>

<221>MOD_RES

<222>(25)..(25)

<223>Amidation

<400>120

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

20 25

<210>121

<211>141

<212>PRT

<213>Homo sapiens

<400>121

Ala Val Ser Glu His Gln Leu Leu His Asp Lys Gly Lys Ser Ile Gln

1 5 10 15

Asp Leu Arg Arg Arg Phe Phe Leu His His Leu Ile Ala Glu Ile His

20 25 30

Thr Ala Glu Ile Arg Ala Thr Ser Glu Val Ser Pro Asn Ser Lys Pro

35 40 45

Ser Pro Asn Thr Lys Asn His Pro Val Arg Phe Gly Ser Asp Asp Glu

50 55 60

Gly Arg Tyr Leu Thr Gln Glu Thr Asn Lys Val Glu Thr Tyr Lys Glu

65 70 75 80

Gln Pro Leu Lys Thr Pro Gly Lys Lys Lys Lys Gly Lys Pro Gly Lys

85 90 95

Arg Lys Glu Gln Glu Lys Lys Lys Arg Arg Thr Arg Ser Ala Trp Leu

100 105 110

Asp Ser Gly Val Thr Gly Ser Gly Leu Glu Gly Asp His Leu Ser Asp

115 120 125

Thr Ser Thr Thr Ser Leu Glu Leu Asp Ser Arg Arg His

130 135 140

<210>122

<211>20

<212>PRT

<213>artificial sequence

<220>

<223>artificial random spiral

<400>122

Gly Gly Pro Gly Gly Pro Gly Pro Gly Gly Pro Gly Gly Pro Gly Pro

1 5 10 15

Gly Gly Pro Gly

twenty

<---

Claims (15)

1. A method of treating hypoparathyroidism in a mammalian patient comprising the step of administering a pharmaceutical composition containing at least one controlled release PTH compound no more frequently than once every 24 hours with a dose of the controlled release PTH compound that corresponds to no more than 40% molar equivalent dose of PTH 1-84 SEQ ID NO: 1 administered subcutaneously at the same dosing frequency required to maintain an albumin-adjusted serum calcium level above 8.5 mg/dl and below 10.5 mg/dl over a 24-hour period, and where soluble in water, the controlled release PTH compound is a compound of formula (IIf-i):

where the unmarked dotted line indicates the attachment to the nitrogen atom of the PTH component having the sequence of SEQ ID NO: 51, by forming an amide bond; and the dotted line marked with an asterisk means the connection of the component:

where m and p independently of each other are an integer in the range and inclusive of 400 to 500. 2. The method of claim 1 wherein the pharmaceutical composition is administered once every 24 hours. 3. The method of claim 1 wherein the pharmaceutical composition is administered once every 48 hours. 4. The method of claim 1 wherein the pharmaceutical composition is administered once every week. 5. The method of claim 1, wherein the dose of the controlled release PTH compound is not more than 30% of the molar equivalent dose of PTH 1-84. 6. The method of claim 1, wherein the pharmaceutical composition is administered via a pen. 7. The method according to p. 1, where the pharmaceutical composition has a pH value in the range and including pH 3 - pH 8. 8. The method of claim 1 wherein the mammalian patient is a human patient. 9. The method according to claim 1, where m and p are about 450.