Patent Description:
Hypoparathyroidism is a rare endocrine disorder of calcium and phosphate metabolism that most often arises as a result of parathyroid gland damage or removal during surgery of the thyroid gland. Hypoparathyroidism is unusual among endocrine disorders in that it has not been treated, until recently, by replacement with the missing hormone, parathyroid hormone, or PTH. Conventional therapy for hypoparathyroidism involves large doses of vitamin D and oral calcium supplementation, which, although often effective, is associated with marked swings in blood Ca<NUM>+ resulting in hypercalcemia and hypocalcemia, excess urinary calcium excretion, nephrocalcinosis, and ectopic calcifications, including vascular, basal ganglia, and lens of eye.

Calcium is the most abundant mineral in the human body, and its tight regulation is required for many critical biological functions, such as bone mineralization, muscle contraction, nerve conduction, hormone release, and blood coagulation. It is particularly important to maintain calcium concentration as stable as possible, because of the high sensitivity of a variety of cell systems or organs, including the central nervous system, muscle, and exo/endocrine glands, to small variations in Ca<NUM>+. PTH is a major regulator of calcium homeostasis.

The inappropriately low or frankly low PTH levels in relation to serum Ca<NUM>+ concentration, characteristic of hypoparathyroidism, leads to decreased renal tubular reabsorption of Ca<NUM>+ and simultaneously, to increased renal tubular reabsorption of phosphate. Thus, the main biochemical abnormalities of hypoparathyroidism are hypocalcemia and hyperphosphatemia. Clinical features of the disease include symptoms of hypocalcemia, such as perioral numbness, paresthesias, and carpal/pedal muscle spasms. Laryngeal spasm, tetany, and seizures are serious and potentially life-threatening complications. Hyperphosphatemia and an elevated calcium x phosphate product contributes to ectopic deposition of insoluble calcium phosphate complexes in soft tissues, including vasculature, brain, kidneys, and other organs.

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

Several concerns arise with prolonged use of calcium and active vitamin D in large doses, particularly with regard to hypercalciuria, kidney stones, nephrocalcinosis and ectopic soft tissue calcification. In addition, conventional therapy with calcium and active vitamin D does not alleviate quality of life complaints nor does it reverse abnormalities in bone remodeling characteristic of the disease. In short, there is a high need for improved therapies for hypoparathyroidism.

In <NUM>, Natpara, PTH(<NUM>-<NUM>), was approved for once-daily subcutaneous injection as an adjunct to vitamin D and calcium in patients with hypoparathyroidism. Natpara, PTH(<NUM>-<NUM>), was approved to control hypocalcemia based on a pivotal trial demonstrating that <NUM> percent of PTH(<NUM>-<NUM>) treated participants achieved normal blood calcium levels on reduced doses of calcium supplements and active forms of vitamin D, compared to <NUM> percent of placebo-treated participants. Following a time course in which serum calcium was monitored after injection, <NUM> percent of patients treated with PTH(<NUM>-<NUM>) developed hypercalcemia at one or more measurements during a <NUM>-hour period. PTH(<NUM>-<NUM>) reduced urinary calcium excretion <NUM>-<NUM> hours after injection but over the <NUM>-hour period, urinary calcium excretion did not change. Similarly, urinary phosphate excretion increased only during the first <NUM> hours after PTH(<NUM>-<NUM>) injection.

While this represents an important advance in the treatment of the disease, Natpara has not demonstrated an ability to reduce incidences of hypercalcemia (elevated serum calcium levels), hypocalcemia (low serum calcium), or hypercalciuria (elevated urinary calcium) relative to conventional therapy in treated patients.

As such, there is a high need for improved PTH based therapies for hypoparathyroidism.

PTH(<NUM>-<NUM>), or teriparatide, was approved by the FDA in <NUM> for the treatment of osteoporosis. Despite not being approved for this indication, PTH(<NUM>-<NUM>) has historically been used for treatment of hypoparathyroidism with patients receiving twice- or thrice-daily injections. To facilitate more physiological PTH levels, clinical studies have been conducted with PTH(<NUM>-<NUM>) administered by pump delivery in comparison with twice-daily injections. Over <NUM>-months, pump delivery produced normal, steady state calcium levels with minimal fluctuation and avoided the rise in serum and urine calcium levels that are evident soon after PTH injection. The marked reduction in urinary calcium excretion when PTH(<NUM>-<NUM>) is administered by pump may indicate that PTH must be continuously exposed to the renal tubule for the renal calcium-conserving effects to be realized. Pump delivery of PTH(<NUM>-<NUM>) achieved simultaneous normalization of markers of bone turnover, serum calcium, and urine calcium excretion. These results were achieved with a <NUM> percent lower daily PTH(<NUM>-<NUM>) dose and a reduced need for magnesium supplementation compared with the twice daily PTH(<NUM>-<NUM>) injection regimen.

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

Long-term daily administration of PTH is associated with a progressive cortical bone loss due to increased bone metabolism. In a <NUM>-year follow-up of patients treated with PTH(<NUM>-<NUM>) (Rubin, JCEM <NUM>) bone turnover markers remained greater than pretreatment values, peaking at the early years after PTH(<NUM>-<NUM>) initiation and declining thereafter but remaining significantly higher than baseline values by year <NUM>. BMD by DXA was consistent with known site-specific effects of PTH, namely increases in lumbar spine and declines in distal <NUM>/<NUM> radius. The decrease observed at the distal one third radius is consistent with the known effects of intermittent PTH to increase cortical porosity and endosteal resorption.

It is an object of this invention to provide for a method of intermittently administering PTH, with improved control of serum and urine calcium, serum phosphorus, and lower elevation of bone turnover markers than currently applied PTH therapies. Preferably intermittent means with daily intervals, or more preferred with weekly intervals.

In the preclinical development program of both Forteo, PTH(<NUM>-<NUM>), and Natpara, PTH(<NUM>-<NUM>), a dose dependent increase in osteosarcoma rate was observed in rats treated with daily injections of the PTH compound. In the Natpara study, dosing of the high dose rats were discontinued due to excessive deaths in this group, primarily from metastatic osteosarcoma. This is felt to be due to the sensitivity of rats to the anabolic effects of intermittent PTH. In contrast, continuous exposure to PTH is known to lack significant bone anabolic activity. As such is an object of this invention to provide for an intermittent PTH replacement therapy that provides for an infusion-like profile of PTH, resulting in improved symptom control with a lower administered dose. Preferably intermittent means with daily intervals, or more preferred with weekly intervals.

<CIT> discloses PTH-containing compounds for use in the treatment of hypoparathyroidism, which may be formulated for sustained release.

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

It is therefore an object of the present invention to at least partially overcome the shortcomings described above.

This object is achieved with a pharmaceutical composition comprising a controlled-release PTH compound or a pharmaceutically acceptable salt, hydrate, or solvate thereof, for use in a method of treating hypoparathyroidism as described in the claims.

It was surprisingly found that such controlled-release PTH compound has a higher potency than PTH <NUM>-<NUM>, so dose adjustment for a pharmaceutical composition comprising such controlled-release PTH compounds can be performed in smaller increments than for pharmaceutical compositions comprising PTH <NUM>-<NUM>, in which doses in response to hypocalcemia or hypercalcemia are doubled or halved, respectively, in order to achieve the right dose. The typical starting dose is <NUM>µg/day, which is titrated down to <NUM>µg/day (<NUM>%) in the event of hypercalcemia, or titrated up to <NUM>µg/day at first (<NUM>%) and then <NUM>µg/day (<NUM>%) in the event of hypocalcemia, with most subjects requiring either <NUM> or <NUM>µg/day.

Within the present invention the terms are used having the meaning as follows.

As used herein the term "increment" refers to the increase or decrease in the amount of controlled-release PTH compound based on a previously administered dose of the same controlled-release PTH compound which increase or decrease is given as a certain percentage based on the weight amount of the controlled-release PTH compound.

As used herein the term "controlled-release PTH compound" refers to any compound, conjugate, crystal or admixture that comprises at least one PTH molecule or moiety and from which the at least one PTH molecule or moiety is released with a release half-life of at least <NUM> hours.

As used herein the terms "release half-life" and "half-life" refer to the time required under physiological conditions (i.e. aqueous buffer, pH <NUM>, <NUM>) until half of all PTH or PTH moieties, respectively, comprised in a controlled-release PTH compound are released from said controlled-release PTH compound.

As used herein the term "PTH" refers to all PTH polypeptides, preferably from mammalian species, more preferably from human and mammalian species, more preferably from human and murine species, as well as their variants, analogs, orthologs, homologs, and derivatives and fragments thereof, that are characterized by raising serum calcium and renal phosphorus excretion, and lowering serum phosphorus and renal calcium excretion. The term "PTH" also refers to all PTHrP polypeptides, such as the polypeptide of SEQ ID NO:<NUM>, that bind to and activate the common PTH/PTHrP1 receptor. Preferably, the term "PTH" refers to the PTH polypeptide of SEQ ID NO:<NUM> as well as its variants, homologs and derivatives exhibiting essentially the same biological activity, i.e. raising serum calcium and renal phosphorus excretion, and lowering serum phosphorus and renal calcium excretion.

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

More preferably, the term "PTH" refers to the sequence of SEQ ID:NOs <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM> and <NUM>. Even more preferably, the term "PTH" refers to the sequence of SEQ ID:NOs <NUM>, <NUM>, <NUM>, <NUM>, <NUM> and <NUM>. In a particularly preferred embodiment the term "PTH" refers to the sequence of SEQ ID NO:<NUM>.

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 reference is a PTH polypeptide sequence and has the sequence of SEQ ID NO:<NUM>. Generally, differences are limited so that the amino acid sequence of the reference and the variant are closely similar overall and, in many regions, identical. Preferably, PTH polypeptide variants are at least <NUM>%, <NUM>%, <NUM>%, or <NUM>% identical to a reference PTH or PTHrP polypeptide, preferably to the PTH polypeptide of SEQ ID NO:<NUM>. By a polypeptide having an amino acid sequence at least, for example, <NUM>% "identical" to a query amino acid sequence, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each <NUM> amino acids of the query amino acid sequence. These alterations of the reference sequence may occur at the amino (N-terminal) or carboxy terminal (C-terminal) positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence. The query sequence may be an entire amino acid sequence of the reference sequence or any fragment specified as described herein. Preferably, the query sequence is the sequence of SEQ ID NO:<NUM>.

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

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

It is also recognized by one of ordinary skill in the art that some amino acid sequences of PTH or PTHrP polypeptides can be varied without significant effect of the structure or function of the polypeptide. Such mutants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as to have little effect on activity. For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in<NPL>, wherein the authors indicate that there are two main approaches for studying the tolerance of the amino acid sequence to change.

The term "PTH polypeptide" also encompasses all PTH and PTHrP polypeptides encoded by PTH and PTHrP analogs, orthologs, and/or species homologs. It is also recognized by one of ordinary skill in the art that PTHrP and PTHrP analogs bind to an activate the common PTH/PTHrP1 receptor, so the term PTH polypeptide also encompasses all PTHrP analogs. As used herein, the term "PTH analog" refers to PTH and PTHrP of different and unrelated organisms which perform the same functions in each organism but which did not originate from an ancestral structure that the organisms' ancestors had in common. Instead, analogous PTH and PTHrP arose separately and then later evolved to perform the same or similar functions. In other words, analogous PTH and PTHrP polypeptides are polypeptides with quite different amino acid sequences but that perform the same biological activity, namely raising serum calcium and renal phosphorus excretion, and lowering serum phosphorus and renal calcium excretion.

As used herein the term "PTH ortholog" refers to PTH and PTHrP within two different species which sequences are related to each other via a common homologous PTH or PTHrP in an ancestral species, but which have evolved to become different from each other.

As used herein, the term "PTH homolog" refers to PTH and PTHrP of different organisms which perform the same functions in each organism and which originate from an ancestral structure that the organisms' ancestors had in common. In other words, homologous PTH polypeptides are polypeptides with quite similar amino acid sequences that perform the same biological activity, namely raising serum calcium and renal phosphorus excretion, and lowering serum phosphorus and renal calcium excretion. Preferably, PTH polypeptide homologs may be defined as polypeptides exhibiting at least <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>% or <NUM>% identity to a reference PTH or PTHrP polypeptide, preferably the PTH polypeptide of SEQ ID NO:<NUM>.

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

As used herein, the term "PTH polypeptide fragment" refers to any polypeptide comprising a contiguous span of a part of the amino acid sequence of a PTH or PTHrP polypeptide, preferably the polypeptide of SEQ ID NO:<NUM>.

More specifically, a PTH polypeptide fragment comprises at least <NUM>, such as at least <NUM>, at least <NUM> or at least <NUM> consecutive amino acids of a PTH or PTHrP polypeptide, more preferably of the polypeptide of SEQ ID NO:<NUM>. A PTH polypeptide fragment may additionally be described as sub-genuses of PTH or PTHrP polypeptides comprising at least <NUM> amino acids, wherein "at least <NUM>" is defined as any integer between <NUM> and the integer representing the C-terminal amino acid of a PTH or PTHrP polypeptide, preferably of the polypeptide of SEQ ID No:<NUM>. Further included are species of PTH or PTHrP polypeptide fragments at least <NUM> amino acids in length, as described above, that are further specified in terms of their N-terminal and C-terminal positions. Also encompassed by the term "PTH polypeptide fragment" as individual species are all PTH or PTHrP polypeptide fragments, at least <NUM> amino acids in length, as described above, that may be particularly specified by a N-terminal and C-terminal position. That is, every combination of a N-terminal and C-terminal position that a fragment at least <NUM> contiguous amino acid residues in length could occupy, on any given amino acid sequence of a PTH or PTHrP polypeptide, preferably the PTH polypeptide of SEQ ID:NO51, is included in the present invention.

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

As the term "PTH" includes the above-described variants, analogs, orthologs, homologs, derivatives and fragments of PTH and PTHrP, all references to specific positions within a reference sequence also include the equivalent positions in variants, analogs, orthologs, homologs, derivatives and fragments of a PTH or PTHrP moiety, even if not specifically mentioned.

As used herein 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 moiety of the surfactant molecules facing the surrounding solvent and the hydrophobic moiety of the surfactant molecule facing inwards, also called "normal-phase micelle". "Invers micelles" have the hydrophilic moiety facing inwards and the hydrophobic moiety facing the surrounding solvent.

As used herein the term "liposome" refers to a vesicle, preferably a spherical vesicle, having at least one lipid bilayer. Preferably, liposomes comprise phospholipids, even more preferably phosphatidylcholine. The term "liposome" refers to various structures and sizes, such as, for example, to multilamellar liposome vesicles (MLV) having more than one concentric lipid bilayer with an average diameter of <NUM> to <NUM>, small unilamellar liposome vesicles (SUV) having one lipid bilayer and an average diameter of <NUM> to <NUM>, large unilamellar liposome vesicles (LUV) having one lipid bilayer and an average diameter of about <NUM> and giant unilamellar vesicles (GUV) having one lipid bilayer and an average diameter of <NUM> to <NUM>. The term "liposome" also includes elastic vesicles such as transferosomes and ethosomes, for example.

As used herein the term "aquasome" refers to spherical nanoparticles having a diameter of <NUM> to <NUM> that comprise at least three layers of self-assembled structure, namely a solid phase nanocrystalline core coated with an oligomeric film to which drug molecules are adsorbed with or without modification of the drug.

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

As used herein the term "LeciPlex" refers to positively charged phospholipid-based vesicular system which comprises soy PC, a cationic agent, and a bio-compatible solvent like PEG <NUM>, PEG <NUM>, diethylene glycol monoethyl ether, tetrahydrofurfuryl alcohol polyethylene glycol ether or <NUM>-pyrrolidoneor N-methyl-<NUM>-pyrrolidone.

As used herein the term "niosome" refers to unilamellar or multilamellar vesicles comprising non-ionic surfactants.

As used herein the term "pharmacosome" refers to ultrafine vesicular, micellar or hexagonal aggregates from lipids covalently bound to biologically active moieties.

As used herein the term "proniosome" refers to dry formulations of surfactant-coated carrier which on rehydration and mild agitation gives niosomes.

As used herein the term "polymersome" refers to an artificial spherical vesicle comprising a membrane formed from amphiphilic synthetic block copolymers and may optionally comprise an aqueous solution in its core. A polymersome has a diameter ranging from <NUM> to <NUM> and larger. The term also includes syntosomes, which are polymersomes engineered to comprise channels that allow certain chemicals to pass through the membrane into or out of the vesicle.

As used herein the term "sphingosome" refers to a concentric, bilayered vesicle in which an aqueous volume is entirely enclosed by a membranous lipid bilayer mainly composed of natural or synthetic sphingolipid.

As used herein the term "transferosome" refers to ultraflexible lipid vesicles comprising an aqueous core that are formed from a mixture of common polar and suitable edge-activated lipids which facilitate the formation of highly curved bilayers which render the transferosome highly deformable.

As used herein the term "ufasome" refers to a vesicle comprising unsaturated fatty acids.

As used herein the term "polypeptide" refers to a peptide comprising up to and including <NUM> amino acid monomers.

As used herein the term "protein" refers to a peptide of more than <NUM> amino acid residues. Preferably a protein comprises at most <NUM> amino acid residues, such as at most <NUM> amino acid residues, such as at most <NUM> amino acid residues, such as at most <NUM> amino acid residues, such as at most <NUM> amino acid residues, such as at most <NUM> amino acid residues, such as at most <NUM> amino acid residues, such as at most <NUM> amino acid residues.

As used herein the term "physiological conditions" refers to an aqueous buffer at pH <NUM>, <NUM>.

As used herein the term "pharmaceutical composition" refers to a composition containing one or more active ingredients, such as for example at least one controlled-release PTH compounds, and one or more excipients, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients of the composition, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing one or more controlled-release PTH compound and a pharmaceutically acceptable excipient.

As used herein the term "liquid composition" refers to a mixture comprising water-soluble controlled-release PTH compound and one or more solvents, such as water.

The term "suspension composition" relates to a mixture comprising water-insoluble controlled-release PTH compound and one or more solvents, such as water.

As used herein, the term "dry composition" means that a pharmaceutical composition is provided in a dry form. Suitable methods for drying are spray-drying and lyophilization, i.e. freeze-drying. Such dry composition of prodrug has a residual water content of a maximum of <NUM> %, preferably less than <NUM>% and more preferably less than <NUM>%, determined according to Karl Fischer. Preferably, the pharmaceutical composition of the present invention is dried by lyophilization.

The term "drug" as used herein refers to a substance used in the treatment, cure, prevention, or diagnosis of a disease or used to otherwise enhance physical or mental well-being. If a drug is conjugated to another moiety, the moiety of the resulting product that originated from the drug is referred to as "biologically active moiety".

As used herein the term "prodrug" refers to a conjugate in which a biologically active moiety is reversibly and covalently connected to a specialized protective group through a reversible linker moiety, also referred to as "reversible prodrug linker moiety", which comprises a reversible linkage with the biologically active moiety and wherein the specialized protective group alters or eliminates undesirable properties in the parent molecule. This also includes the enhancement of desirable properties in the drug and the suppression of undesirable properties. The specialized non-toxic protective group is referred to as "carrier". A 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 comprising a biologically active moiety which is covalently and reversibly conjugated to a carrier moiety via a reversible prodrug linker moiety, which covalent and reversible conjugation of the carrier to the reversible prodrug linker moiety is either directly or through a spacer. Such conjugate releases the formerly conjugated biologically active moiety in the form of a free unmodified drug.

A "biodegradable linkage" or a "reversible linkage" is a linkage that is hydrolytically degradable, i.e. cleavable, in the absence of enzymes under physiological conditions (aqueous buffer at pH <NUM>, <NUM>) with a half-life ranging from one hour to three months, preferably from one hour to two months, even more preferably from one hour to one month, even more preferably from one hour to three weeks, most preferably from one hour to two weeks. Accordingly, a stable linkage is a linkage having a half-life under physiological conditions (aqueous buffer at pH <NUM>, <NUM>) of more than three months.

As used herein, the term "traceless prodrug linker" means a reversible prodrug linker, i.e. a linker moiety reversibly and covalently connecting the biologically active moiety with the carrier, which upon cleavage releases the drug in its free form. As used herein, the term "free form" of a drug means the drug in its unmodified, pharmacologically active form.

As used herein, the term "excipient" refers to a diluent, adjuvant, or vehicle with which the therapeutic, such as a drug or prodrug, is administered. Such pharmaceutical excipient can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, including peanut oil, soybean oil, mineral oil and sesame oil. Water is a preferred excipient when the pharmaceutical composition is administered orally. Saline and aqueous dextrose are preferred excipients when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions are preferably employed 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, dried skim milk, glycerol, propylene, glycol, water and ethanol. The pharmaceutical composition, if desired, can also contain minor amounts of wetting or emulsifying agents, pH buffering agents, like, for example, acetate, succinate, tris, carbonate, phosphate, HEPES (<NUM>-(<NUM>-hydroxyethyl)-<NUM>-piperazineethanesulfonic acid), MES (<NUM>-(A-morpholino)ethanesulfonic acid), or can contain detergents, like Tween, poloxamers, poloxamines, CHAPS, Igepal, or amino acids like, for example, glycine, lysine, or histidine. These pharmaceutical compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders and sustained-release formulations. The pharmaceutical composition can be formulated as a suppository, with traditional binders and excipients such as triglycerides. Oral formulation can include standard excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Such compositions will contain a therapeutically effective amount of the drug or biologically active moiety, together with a suitable amount of excipient so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

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

As used herein, the term "moiety" means a part of a molecule, which lacks one or more atom(s) compared to the corresponding reagent. If, for example, a reagent of the formula "H-X-H" reacts with another reagent and becomes part of the reaction product, the corresponding moiety of the reaction product has the structure "H-X-" or "-X-", whereas each "-" indicates attachment to another moiety. Accordingly, a biologically active moiety is released from a prodrug as a drug.

It is understood that if the sequence or chemical structure of a group of atoms is provided which group of atoms is attached to two moieties or is interrupting a moiety, said sequence or chemical structure can be attached to the two moieties in either orientation, unless explicitly stated otherwise. For example, a moiety "-C(O)N(R<NUM>)-" can be attached to two moieties or interrupting a moiety either as "-C(O)N(R<NUM>)-" or as "-N(R<NUM>)C(O)-". Similarly, a moiety
<CHM>
can be attached to two moieties or can interrupt a moiety either as
<CHM>
or as
<CHM>.

As used herein, the term "functional group" means a group of atoms which can react with other groups of atoms. Functional groups include but are not limited to the following groups: carboxylic acid (-(C=O)OH), primary or secondary amine (-NH<NUM>, -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 (>N-N<), 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 of the present invention comprise one or more acidic or basic groups, the invention also comprises their corresponding pharmaceutically or toxicologically acceptable salts, in particular their pharmaceutically utilizable salts. Thus, the controlled-release PTH compound of the present invention comprising acidic groups can be used according to the invention, for example, as alkali metal salts, alkaline earth metal salts or as ammonium salts. More precise examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts or salts with ammonia or organic amines such as, for example, ethylamine, ethanolamine, triethanolamine or amino acids. Controlled-release PTH compound of the present invention comprising one or more basic groups, i.e. groups which can be protonated, can be present and can be used according to the invention in the form of their addition salts with inorganic or organic acids. Examples for suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acids, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfaminic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to the person skilled in the art. For the person skilled in the art further methods are known for converting the basic group into a cation like the alkylation of an amine group resulting in a positively-charge ammonium group and an appropriate counterion of the salt. If the controlled-release PTH compound of the present invention simultaneously comprise acidic and basic groups, the invention also includes, in addition to the salt forms mentioned, inner salts or betaines (zwitterions). The respective salts can be obtained by customary methods which are known to the person skilled in the art like, 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 of the present invention which, owing to low physiological compatibility, are not directly suitable for use in pharmaceuticals 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 cause harm when administered to a patient and preferably means approved by a regulatory agency, such as the EMA (Europe) and/or the FDA (US) and/or any other national regulatory agency for use in animals, preferably for use in humans.

As used herein the term "about" in combination with a numerical value is used to indicate a range ranging from and including the numerical value plus and minus no more than <NUM>% of said numerical value, more preferably no more than <NUM>% of said numerical value, even more preferably no more than <NUM>% of said numerical value and most preferably no more than <NUM>% of said numerical value. For example, the phrase "about <NUM>" is used to mean a range ranging from and including <NUM> +/- <NUM>%, i.e. ranging from and including <NUM> to <NUM>; preferably <NUM> +/-<NUM>%, i.e. ranging from and including <NUM> to <NUM>; even more preferably ranging from and including <NUM> +/-<NUM>%, i.e. ranging from and including <NUM> to <NUM>; and most preferably <NUM> +/-<NUM>%, i.e. ranging from and including <NUM> to <NUM>. It is understood that a percentage given as "about <NUM>%" does not mean "<NUM>% +/- <NUM>%", i.e. ranging from and including <NUM> to <NUM>%, but "about <NUM>%" means ranging from and including <NUM> to <NUM>%, i.e. plus and minus <NUM>% of the numerical value which is <NUM>.

As used herein, the term "polymer" means a molecule comprising repeating structural units, i.e. the monomers, connected by chemical bonds in a linear, circular, branched, crosslinked or dendrimeric way or a combination thereof, which may be of synthetic or biological origin or a combination of both. It is understood that a polymer may also comprise one or more other chemical groups and/or moieties, such as, for example, one or more functional groups. Preferably, a soluble polymer has a molecular weight of at least <NUM> kDa, e.g. a molecular weight of at least <NUM> kDa, a molecular weight of at least <NUM> kDa, a molecular weight of at least <NUM> kDa or a molecular weight of at least <NUM> kDa. If the polymer is soluble, it preferable has a molecular weight of at most <NUM> kDa, such as at most <NUM> kDa, such as at most <NUM> kDa, such as at most <NUM> kDa, such as at most <NUM> kDa, such as at most <NUM> kDa. It is understood that for insoluble polymers, such as hydrogels, no meaningful molecular weight ranges can be provided. It is understood that also a protein is a polymer in which the amino acids are the repeating structural units, even though the side chains of each amino acid may be different.

As used herein, the term "polymeric" means a reagent or a moiety comprising one or more polymers or polymer moieties. A polymeric reagent or moiety may optionally also comprise one or more other moiety/moieties, which are preferably selected from the group consisting of:.

The person skilled in the art understands that the polymerization products obtained from a polymerization reaction do not all have the same molecular weight, but rather exhibit a molecular weight distribution. Consequently, the molecular weight ranges, molecular weights, ranges of numbers of monomers in a polymer and numbers of monomers in a polymer as used herein, refer to the number average molecular weight and number average of monomers, i.e. to the arithmetic mean of the molecular weight of the polymer or polymeric moiety and the arithmetic mean of the number of monomers of the polymer or polymeric moiety.

Accordingly, in a polymeric moiety comprising "x" monomer units any integer given for "x" therefore corresponds to the arithmetic mean number of monomers. Any range of integers given for "x" provides the range of integers in which the arithmetic mean numbers of monomers lies. An integer for "x" given as "about x" means that the arithmetic mean numbers of monomers lies in a range of integers of x +/- <NUM>%, preferably x +/- <NUM>%, more preferably x +/- <NUM>% and most preferably x +/- <NUM>%.

As used herein, the term "number average molecular weight" means the ordinary arithmetic mean of the molecular weights of the individual polymers.

As used herein the term "water-soluble" with reference to a carrier means that when such carrier is part of the controlled-release PTH compound of the present invention at least <NUM> of the controlled-release PTH compound comprising such water-soluble carrier can be dissolved in one liter of water at <NUM> to form a homogeneous solution. Accordingly, the term "water-insoluble" with reference to a carrier means that when such carrier is part of a controlled-release PTH compound of the present invention less than <NUM> of the controlled-release PTH compound comprising such water-insoluble carrier can be dissolved in one liter of water at <NUM> to form a homogeneous solution.

As used herein, the term "hydrogel" means a hydrophilic or amphiphilic polymeric network composed of homopolymers or copolymers, which is insoluble due to the presence of covalent chemical crosslinks. The crosslinks provide the network structure and physical integrity.

As used herein the term "thermogelling" means a compound that is a liquid or a low viscosity solution having a viscosity of less than <NUM> cps at <NUM> at a shear rate of about <NUM> /second at a low temperature, which low temperature ranges between about <NUM> to about <NUM>, but which is a higher viscosity compound of less than <NUM> cps at <NUM> at a shear rate of about <NUM>/second at a higher temperature, which higher temperature ranges between about <NUM> to about <NUM>, such as at about <NUM>.

As used herein, the term "PEG-based" in relation to a moiety or reagent means that said moiety or reagent comprises PEG. Preferably, a PEG-based moiety or reagent comprises at least <NUM>% (w/w) PEG, such as at least <NUM>% (w/w) PEG, such as at least <NUM>% (w/w) PEG, such as at least <NUM>% (w/w) PEG, such as at least <NUM>% (w/w), such as at least <NUM> (w/w) PEG, such as at least <NUM>% (w/w) PEG, such as at least <NUM>% (w/w) PEG, such as at least <NUM>% (w/w) PEG, such as at least <NUM>%. The remaining weight percentage of the PEG-based moiety or reagent are other moieties preferably selected from the following moieties and linkages:.

As used herein, the term "PEG-based comprising at least X% PEG" in relation to a moiety or reagent means that said moiety or reagent comprises at least X% (w/w) ethylene glycol units (-CH<NUM>CH<NUM>O-), wherein the ethylene glycol units may be arranged blockwise, alternating or may be randomly distributed within the moiety or reagent and preferably all ethylene glycol units of said moiety or reagent are present in one block; the remaining weight percentage of the PEG-based moiety or reagent are other moieties preferably selected from the following moieties and linkages:.

The term "hyaluronic acid-based comprising at least X% hyaluronic acid" is used accordingly.

The term "substituted" as used herein means that one or more -H atom(s) of a molecule or moiety are replaced by a different atom or a group of atoms, which are referred to as "substituent".

Preferably, the one or more further optional substituents are independently of each other selected from the group consisting of halogen, -CN, -COORx1, -ORx1, -C(O)Rx1, -C(O)N(Rx1Rx1a), -S(O)<NUM>N(Rx1Rx1a), -S(O)N(Rx1Rx1a), -S(O)<NUM>Rx1, -S(O)Rx1, -N(Rx1)S(O)<NUM>N(Rx1aRx1b), -SRx1, -N(Rx1Rx1a), -NO<NUM>, -OC(O)Rx1, -N(Rx1)C(O)Rx1a, -N(Rx1)S(O)<NUM>Rx1a, -N(Rx1)S(O)Rx1a, -N(Rx1)C(O)ORx1a, -N(Rx1)C(O)N(Rx1aRx1b), -OC(O)N(Rx1Rx1a), -T<NUM>, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, and C<NUM>-<NUM> alkynyl; wherein -T<NUM>, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, and C<NUM>-<NUM> alkynyl are optionally substituted with one or more -Rx2, which are the same or different and wherein C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, and C<NUM>-<NUM> alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T<NUM>-, -C(O)O-, -O-, -C(O)-, -C(O)N(Rx3)-, -S(O)<NUM>N(Rx3)-, -S(O)N(Rx3)-, -S(O)<NUM>-, -S(O)-, -N(Rx3)S(O)<NUM>N(Rx3a)-, -S-, -N(Rx3)-, -OC(ORx3)(Rx3a)-, -N(Rx3)C(O)N(Rx3a)-, and -OC(O)N(Rx3)-;.

More preferably, the one or more further optional substituents are independently of each other selected from the group consisting of halogen, -CN, -COORx1, -ORx1, -C(O)Rx1, -C(O)N(Rx1Rx1a), -S(O)<NUM>N(Rx1Rx1a), -S(O)N(Rx1Rx1a), -S(O)<NUM>Rx1, -S(O)Rx1, -N(Rx1)S(O)<NUM>N(Rx1aRx1b), -SRx1, -N(Rx1Rx1a), -NO<NUM>, -OC(O)Rx1, -N(Rx1)C(O)Rx1a, -N(Rx1)S(O)<NUM>Rx1a, -N(Rx1)S(O)Rx1a, -N(Rx1)C(O)ORx1a, -N(Rx1)C(O)N(Rx1aRx1b), -OC(O)N(Rx1Rx1a), -T<NUM>, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, and C<NUM>-<NUM> alkynyl; wherein -T<NUM>, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, and C<NUM>-<NUM> alkynyl are optionally substituted with one or more -Rx2, which are the same or different and wherein C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, and C<NUM>-<NUM> alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T<NUM>-, -C(O)O-, -O-, -C(O)-, -C(O)N(Rx3)-, -S(O)<NUM>N(Rx3)-, -S(O)N(Rx3)-, -S(O)<NUM>-, -S(O)-, -N(Rx3)S(O)<NUM>N(Rx3a)-, -S-, -N(Rx3)-, -OC(ORx3)(Rx3a)-, -N(Rx3)C(O)N(Rx3a)-, and -OC(O)N(Rx3)-;.

Preferably, a maximum of <NUM> -H atoms of an optionally substituted molecule are independently replaced by a substituent, e.g. <NUM> -H atoms are independently replaced by a substituent, <NUM> -H atoms are independently replaced by a substituent, <NUM> -H atoms are independently replaced by a substituent, <NUM> -H atoms are independently replaced by a substituent, or <NUM> -H atom is replaced by a substituent.

The term "interrupted" means that a moiety is inserted between two carbon atoms or - if the insertion is at one of the moiety's ends - between a carbon or heteroatom and a hydrogen atom, preferably between a carbon and a hydrogen atom.

As used herein, the term "C<NUM>-<NUM> alkyl" alone or in combination means a straight-chain or branched alkyl moiety having <NUM> to <NUM> carbon atoms. If present at the end of a molecule, examples of straight-chain or branched C<NUM>-<NUM> alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. When two moieties of a molecule are linked by the C<NUM>-<NUM> alkyl, then examples for such C<NUM>-<NUM> alkyl groups are -CH<NUM>-, -CH<NUM>-CH<NUM>-, -CH(CH<NUM>)-, -CH<NUM>-CH<NUM>-CH<NUM>-, -CH(C<NUM>H<NUM>)-, -C(CH<NUM>)<NUM>-. Each hydrogen of a C<NUM>-<NUM> alkyl carbon may optionally be replaced by a substituent as defined above. Optionally, a C<NUM>-<NUM> alkyl may be interrupted by one or more moieties as defined below.

As used herein, the term "C<NUM>-<NUM> alkyl" alone or in combination means a straight-chain or branched alkyl moiety having <NUM> to <NUM> carbon atoms. If present at the end of a molecule, examples of straight-chain and branched C<NUM>-<NUM> alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, <NUM>-methylbutyl, <NUM>,<NUM>-dimethylpropyl, n-hexyl, <NUM>-methylpentyl, <NUM>-methylpentyl, <NUM>,<NUM>-dimethylbutyl, <NUM>,<NUM>-dimethylbutyl and <NUM>,<NUM>-dimethylpropyl. When two moieties of a molecule are linked by the C<NUM>-<NUM> alkyl group, then examples for such C<NUM>-<NUM> alkyl groups are -CH<NUM>-, -CH<NUM>-CH<NUM>-, -CH(CH<NUM>)-, -CH<NUM>-CH<NUM>-CH<NUM>-, -CH(C<NUM>H<NUM>)- and -C(CH<NUM>)<NUM>-. Each hydrogen atom of a C<NUM>-<NUM> carbon may optionally be replaced by a substituent as defined above. Optionally, a C<NUM>-<NUM> alkyl may be interrupted by one or more moieties as defined below.

Accordingly, "C<NUM>-<NUM> alkyl", "C<NUM>-<NUM> alkyl" or "C<NUM>-<NUM> alkyl" means an alkyl chain having <NUM> to <NUM>, <NUM> to <NUM> or <NUM> to <NUM> carbon atoms, respectively, wherein each hydrogen atom of the C<NUM>-<NUM>, C<NUM>-<NUM> or C<NUM>-<NUM> carbon may optionally be replaced by a substituent as defined above. Optionally, a C<NUM>-<NUM> or C<NUM>-<NUM> alkyl may be interrupted by one or more moieties as defined below.

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

Accordingly, the term "C<NUM>-<NUM> alkenyl", "C<NUM>-<NUM> alkenyl" or "C<NUM>-<NUM> alkenyl" alone or in combination means a straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon double bond having <NUM> to <NUM>, <NUM> to <NUM> or <NUM> to <NUM> carbon atoms. Each hydrogen atom of a C<NUM>-<NUM> alkenyl, C<NUM>-<NUM> alkenyl or C<NUM>-<NUM> alkenyl group may optionally be replaced by a substituent as defined above. Optionally, a C<NUM>-<NUM> alkenyl, C<NUM>-<NUM> alkenyl or C<NUM>-<NUM> alkenyl may be interrupted by one or more moieties as defined below.

As used herein, the term "C<NUM>-<NUM> alkynyl" alone or in combination means straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon triple bond having <NUM> to <NUM> carbon atoms. If present at the end of a molecule, examples are -C=CH, -CH<NUM>-C≡CH, CH<NUM>-CH<NUM>-C=CH and CH<NUM>-C≡C-CH<NUM>. When two moieties of a molecule are linked by the alkynyl group, then an example is -C=C-. Each hydrogen atom of a C<NUM>-<NUM> alkynyl group may optionally be replaced by a substituent as defined above. Optionally, one or more double bond(s) may occur. Optionally, a C<NUM>-<NUM> alkynyl may be interrupted by one or more moieties as defined below.

Accordingly, as used herein, the term "C<NUM>-<NUM> alkynyl", "C<NUM>-<NUM> alkynyl" and "C<NUM>-<NUM> alkynyl" alone or in combination means a straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon triple bond having <NUM> to <NUM>, <NUM> to <NUM> or <NUM> to <NUM> carbon atoms, respectively. Each hydrogen atom of a C<NUM>-<NUM> alkynyl, C<NUM>-<NUM> alkynyl or C<NUM>-<NUM> alkynyl group may optionally be replaced by a substituent as defined above. Optionally, one or more double bond(s) may occur.

Optionally, a C<NUM>-<NUM> alkynyl, C<NUM>-<NUM> alkynyl or C<NUM>-<NUM> alkynyl may be interrupted by one or more moieties as defined below.

As mentioned above, a C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkenyl, C<NUM>-<NUM> alkenyl, C<NUM>-<NUM> alkenyl, C<NUM>-<NUM> alkenyl, C<NUM>-<NUM> alkynyl, C<NUM>-<NUM> alkynyl, C<NUM>-<NUM> alkenyl or C<NUM>-<NUM> alkynyl may optionally be interrupted by one or more moieties which are preferably selected from the group consisting of
<CHM>
<CHM>
wherein dashed lines indicate attachment to the remainder of the moiety or reagent; and -R and -Ra are independently of each other selected from the group consisting of -H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

As used herein, the term "C<NUM>-<NUM> cycloalkyl" means a cyclic alkyl chain having <NUM> to <NUM> carbon atoms, which may be saturated or unsaturated, e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl. Each hydrogen atom of a C<NUM>-<NUM> cycloalkyl carbon may be replaced by a substituent as defined above. The term "C<NUM>-<NUM> cycloalkyl" also includes bridged bicycles like norbornane or norbornene.

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

As used herein, the term "<NUM>- to <NUM>-membered heterocyclyl" or "<NUM>- to <NUM>-membered heterocycle" means a ring with <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM> or <NUM> ring atoms that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or un-saturated) wherein at least one ring atom up to <NUM> ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)<NUM>-), oxygen and nitrogen (including =N(O)-) and wherein the ring is linked to the rest of the molecule via a carbon or nitrogen atom. Examples for <NUM>- to <NUM>-membered heterocycles include but are not limited to aziridine, oxirane, thiirane, azirine, oxirene, thiirene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazoline, thiadiazole, thiadiazoline, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine and homopiperazine. Each hydrogen atom of a <NUM>- to <NUM>-membered heterocyclyl or <NUM>- to <NUM>-membered heterocyclic group may be replaced by a substituent as defined below.

As used herein, the term "<NUM>- to <NUM>-membered heterobicyclyl" or "<NUM>- to <NUM>-membered heterobicycle" means a heterocyclic moiety of two rings with <NUM> to <NUM> ring atoms, where at least one ring atom is shared by both rings and that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or un-saturated) wherein at least one ring atom up to <NUM> ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)<NUM>-), oxygen and nitrogen (including =N(O)-) and wherein the ring is linked to the rest of the molecule via a carbon or nitrogen atom. Examples for an <NUM>- to <NUM>-membered heterobicycle are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine and pteridine. The term <NUM>- to <NUM>-membered heterobicycle also includes spiro structures of two rings like <NUM>,<NUM>-dioxa-<NUM>-azaspiro[<NUM>]decane or bridged heterocycles like <NUM>-aza-bicyclo[<NUM>. <NUM>]octane. Each hydrogen atom of an <NUM>- to <NUM>-membered heterobicyclyl or <NUM>- to <NUM>-membered heterobicycle carbon may be replaced by a substituent as defined below.

Similary, the term "<NUM>- to <NUM>-membered heteropolycyclyl" or "<NUM>- to <NUM>-membered heteropolycycle" means a heterocyclic moiety of more than two rings with <NUM> to <NUM> ring atoms, preferably of three, four or five rings, where two neighboring rings share at least one ring atom and that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or unsaturated), wherein at least one ring atom up to <NUM> ring atoms are replaced by a heteroatom selected from the group of sulfur (including -S(O)-, -S(O)<NUM>-), oxygen and nitrogen (including =N(O)-) and wherein the ring is linked to the rest of a molecule via a carbon or nitrogen atom.

It is understood that the phrase "the pair Rx/Ry is joined together with the atom to which they are attached to form a C<NUM>-<NUM> cycloalkyl or a <NUM>- to <NUM>-membered heterocyclyl" in relation with a moiety of the structure
<CHM>
means that Rx and Ry form the following structure:
<CHM>
wherein R is C<NUM>-<NUM> cycloalkyl or <NUM>- to <NUM>-membered heterocyclyl.

It is also understood that the phrase "the pair Rx/Ry is joint together with the atoms to which they are attached to form a ring A" in relation with a moiety of the structure
<CHM>
means that Rx and Ry form the following structure:
<CHM>.

As used herein, "halogen" means fluoro, chloro, bromo or iodo. It is generally preferred that halogen is fluoro or chloro.

In general, the term "comprise" or "comprising" also encompasses "consist of" or "consisting of '.

Preferably, the pharmaceutical composition comprising a controlled-release PTH compound is administered in accordance with a dosage regimen in which dose adjustment in response to hypocalcemia or hypercalcemia is performed in increments of no more than <NUM>%, more preferably in increments of no more than <NUM>% and most preferably in increments of no more than <NUM>%.

In on embodiment the pharmaceutical composition comprising a controlled-release PTH compound is administered in accordance with a dosage regimen in which dose adjustment in response to hypocalcemia or hypercalcemia is performed in increments of <NUM>%.

More preferably, the pharmaceutical composition comprising a controlled-release PTH compound is administered in accordance with a dosage regimen in which dose adjustment in response to hypocalcemia or hypercalcemia is performed in increments of <NUM>%.

Even more preferably, the pharmaceutical composition comprising a controlled-release PTH compound is administered in accordance with a dosage regimen in which dose adjustment in response to hypocalcemia or hypercalcemia is performed in increments of <NUM>%.

Most preferably, the pharmaceutical composition comprising a controlled-release PTH compound is administered in accordance with a dosage regimen in which dose adjustment in response to hypocalcemia or hypercalcemia is performed in increments of <NUM>%.

Preferably, the pharmaceutical comprising the controlled-release PTH compound it administered to the patient no more often than once every <NUM> hours, such as every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, every <NUM> hours, once a week, once every two weeks.

In one embodiment the pharmaceutical composition comprising the controlled-release PTH compound is administered every <NUM> hours.

In another embodiment the pharmaceutical composition comprising the controlled-release PTH compound is administered every <NUM> hours.

In another embodiment the pharmaceutical composition comprising the controlled-release PTH compound is administered once every week.

Preferably, the pharmaceutical composition comprising the PTH compound is administered to a patient via subcutaneous administration, preferably via subcutaneous injection.

In one embodiment the pharmaceutical composition for use of the present invention is performed with a syringe. In another embodiment the pharmaceutical composition for use of the present invention is administered with a pen injector. In another embodiment the pharmaceutical composition for use of the present invention is administered with an auto injector.

The controlled-release PTH compound is water-soluble.

Such water-soluble controlled-release PTH compound is a compound of formula (Ia)
<CHM>
wherein.

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

The moiety -L<NUM>- is either conjugated to a functional group of the side chain of an amino acid residue of -D or to the N-terminal amine functional group of -D.

Preferably, the amino acid residue of -D to which -L<NUM>- is conjugated comprises a primary or secondary amine functional group. Most preferably the amino acid residue of -D to which -L<NUM>- is conjugated comprises a primary amine functional group.

In certain embodiments -L<NUM>- is conjugated to a functional group of the side chain of an amino acid residue of -D. Preferably said amino acid is selected from the group consisting of histidine, lysine, tryptophan and arginine. Even more preferably said amino acid is selected from the group consisting of lysine and arginine.

In one embodiment -L<NUM>- is conjugated to a functional group of the side chain of a histidine of -D.

In another embodiment -L<NUM>- is conjugated to a functional group of the side chain of a lysine of -D.

In another embodiment -L<NUM>- is conjugated to a functional group of the side chain of a tryptophan of -D.

In another embodiment -L<NUM>- is conjugated to a functional group of the side chain of an arginine of -D.

In a preferred embodiment -L<NUM>- is conjugated to the N-terminal amine functional group of -D. The moiety -L<NUM>-L<NUM>- is
<CHM>
wherein.

The controlled-release PTH compound of the present invention is of formula (Ia) with x = <NUM>.

-Z comprises a moiety of formula (b)
<CHM>
wherein.

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

Most preferably m and p of formula (b) are about <NUM>.

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

In a preferred embodiment the PTH compound in the pharmaceutical composition of the present invention is of formula (IIf-i):
<CHM>
wherein.

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

In a preferred embodiment the residual activity of the controlled-release PTH compound in the form of a PTH prodrug is less than <NUM>%, more preferably less than <NUM>%, even more preferably less than <NUM>%, even more preferably less than <NUM>%, even more preferably less than <NUM>% and most preferably less than <NUM>%.

As used herein the term "residual activity" refers to the activity exhibited by the PTH prodrug with the PTH moiety bound to a carrier in relation to the activity exhibited by the corresponding free PTH. In this context the term "activity" refers to binding to an activation of the PTH/PTHrP <NUM> receptor resulting in activation of adenylate cyclase to generate cAMP, phospholipase C to generate intracellular calcium, or osteoblastic expression of RANKL (which binds to RANK (Receptor Activator of Nuclear Factor kB) on osteoclasts. It is understood that measuring the residual activity of the PTH prodrug of the present invention takes time during which a certain amount of PTH will be released from the PTH prodrug of the present invention and that such released PTH will distort the results measured for the PTH prodrug. It is thus accepted practice to test the residual activity of a prodrug with a conjugate in which the drug moiety, in this case PTH, is non-reversibly, i.e. stably, bound to a carrier, which as closely as possible resembles the structure of the PTH prodrug for which residual activity is to be measured.

Preferably, the pharmaceutical composition comprising at least one controlled-release PTH compound of the present invention has a pH ranging from and including pH <NUM> to pH <NUM>. More preferably, the pharmaceutical composition has a pH ranging from and including pH <NUM> to pH <NUM>. Most preferably, the pharmaceutical composition has a pH ranging from and including pH <NUM> to pH <NUM>.

In one embodiment the pharmaceutical composition comprising at least one controlled-release PTH compound of the present invention is a liquid or suspension formulation. It is understood that the pharmaceutical composition is a suspension formulation if the controlled-release PTH compound of the present invention is water-insoluble.

In another embodiment the pharmaceutical composition comprising at least one controlled-release PTH compound of the present invention is a dry formulation which is reconstituted before administration to a patient.

Such liquid, suspension, dry or reconstituted pharmaceutical composition comprises at least one excipient. Excipients used in parenteral formulations may be categorized as, for example, buffering agents, isotonicity modifiers, preservatives, stabilizers, anti-adsorption agents, oxidation protection agents, viscosifiers/viscosity enhancing agents, or other auxiliary agents. However, in some cases, one excipient may have dual or triple functions. Preferably, the at least one excipient comprised in the pharmaceutical composition of the present invention is selected from the group consisting of.

Preferably, the patient treated for hypoparathyroidism of the present invention is a mammalian patient, preferably a human patient.

Side chain protected PTH(<NUM>-<NUM>) (SEQ ID NO:<NUM>) on TCP resin having Boc protected N-terminus and ivDde protected side chain of Lys26 (synthesized by Fmoc-strategy) was obtained from a custom peptide synthesis supplier.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus (synthesized by Fmoc-strategy) was obtained from a custom peptide synthesis supplier.

PEG 2x20 kDa maleimide, Sunbright GL2-400MA was purchased from NOF Europe N. , Grobbendonk, Belgium. S-Trityl-<NUM>-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 a different supplier is mentioned.

Compound 11a (examples <NUM>-<NUM>) was synthesized following the procedure described in patent <CIT>, example <NUM>.

Syringes equipped with polyethylenene frits (MultiSynTech GmbH, Witten, Germany) were used as reaction vessels or for washing steps of peptide resins.

General procedure for the removal of ivDde protecting group from side chain protected PTH on resin: The resin was pre-swollen in DMF for <NUM> and the solvent was discarded. The ivDde group was removed by incubating the resin with DMF/hydrazine hydrate <NUM>/<NUM> (v/v, <NUM>/g resin) for <NUM> x <NUM>. For each step fresh DMF/hydrazine hydrate solution was used. Finally, the resin was washed with DMF (<NUM> x), DCM (<NUM> x) and dried in vacuo.

General procedure for the removal of Fmoc protecting group from protected PTH on resin: The resin was pre-swollen in DMF for <NUM> and the solvent was discarded. The Fmoc group was removed by incubating the resin with DMF/piperidine/DBU <NUM>/<NUM>/<NUM> (v/v/v, <NUM>/g resin) for <NUM> x <NUM>. For each step fresh DMF/piperidine/DBU hsolution was used. Finally, the resin was washed with DMF (<NUM> x), DCM (<NUM> x) and dried in vacuo.

For preparative RP-HPLC a Waters <NUM> controller and a <NUM> Dual Absorbance Detector was used, equipped with the following columns: Waters XBridge™ BEH300 Prep C18 <NUM>, <NUM> x <NUM>, flow rate <NUM>/min, or Waters XBridge™ BEH300 Prep C18 <NUM>, <NUM> x <NUM>, flow rate <NUM>/min. Linear gradients of solvent system A (water containing <NUM> % TFA v/v) and solvent system B (acetonitrile containing <NUM> % TFA v/v) were used. HPLC fractions containing product were pooled and lyophilized if not stated otherwise.

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

Ion exchange chromatography (IEX) was performed using an Amersham Bioscience AEKTAbasic system equipped with a MacroCap SP cation exchanger column (Amersham Bioscience/GE Healthcare). <NUM> acetic acid pH <NUM> (solvent A) and <NUM> acetic acid, <NUM> NaCl, pH <NUM> (solvent B) were used as mobile phases.

Size exclusion chromatography (SEC) was performed using an Amersham Bioscience AEKTAbasic system equipped with HiPrep <NUM>/<NUM> desalting columns (Amersham Bioscience/GE Healthcare). <NUM> % (v/v) acetic acid was used as mobile phase.

Analytical ultra-performance LC (UPLC)-MS was performed on a Waters Acquity system equipped with a Waters BEH300 C18 column (<NUM> × <NUM>, <NUM> particle size, flow: <NUM>/min; solvent A: water containing <NUM>% TFA (v/v), solvent B: acetonitrile containing <NUM>% TFA (v/v)) coupled to a LTQ Orbitrap Discovery mass spectrometer from Thermo Scientific or coupled to a Waters Micromass ZQ.

Linker reagent If was synthesized according to the following scheme:
<CHM>.

To a solution of N-methyl-N-Boc-ethylenediamine (<NUM>, <NUM> mmol) and NaCNBH<NUM> (<NUM>, <NUM> mmol) in MeOH (<NUM>) was added <NUM>,<NUM>,<NUM>-trimethoxybenzaldehyde (<NUM>, <NUM> mmol) portion wise. The mixture was stirred at rt for <NUM>, acidified with <NUM> HCl (<NUM>) and stirred further <NUM>. The reaction mixture was added to saturated NaHCO<NUM> solution (<NUM>) and extracted <NUM> x with DCM. The combined organic phases were dried over Na<NUM>SO<NUM> and the solvents were evaporated in vacuo. The resulting N-methyl-N-Boc-N'-Tmob-ethylenediamine 1a was dried in high vacuum and used in the next reaction step without further purification.

To a solution of 1a (<NUM>, <NUM> mmol) in DCM (<NUM>) COMU (<NUM>, <NUM> mmol), N-Fmoc-N-Me-Asp(OBn)-OH (<NUM>, <NUM> mmol) and <NUM>,<NUM>,<NUM>-collidine (<NUM>, <NUM> mmol) were added. The reaction mixture was stirred for <NUM> at rt, diluted with DCM (<NUM>) and washed <NUM> x with <NUM> H<NUM>SO<NUM> (<NUM>) and <NUM> x with brine (<NUM>). The aqueous phases were re-extracted with DCM (<NUM>). The combined organic phases were dried over Na<NUM>SO<NUM>, filtrated and the residue concentrated to a volume of <NUM>. 1b was purified using flash chromatography.

To a solution of 1b (<NUM>, max. <NUM> mmol ref. to N-Fmoc-N-Me-Asp(OBn)-OH) in THF (<NUM>) DBU (<NUM>, <NUM> % v/v) was added. The solution was stirred for <NUM> at rt, diluted with DCM (<NUM>) and washed <NUM> x with <NUM> H<NUM>SO<NUM> (<NUM>) and <NUM> x with brine (<NUM>). The aqueous phases were re-extracted with DCM (<NUM>). The combined organic phases were dried over Na<NUM>SO<NUM> and filtrated. 1c was isolated upon evaporation of the solvent and used in the next reaction without further purification.

1c (<NUM>, <NUM> mmol, crude) was dissolved in acetonitrile (<NUM>) and COMU (<NUM>, <NUM> mmol), <NUM>-tritylmercaptohexanoic acid (<NUM>, <NUM> mmol) and <NUM>,<NUM>,<NUM>-collidine (<NUM>, <NUM> mmol) were added. The reaction mixture was stirred for <NUM> at rt, diluted with DCM (<NUM>) and washed <NUM> x with <NUM> H<NUM>SO<NUM> (<NUM>) and <NUM> x with brine (<NUM>). The aqueous phases were re-extracted with DCM (<NUM>). The combined organic phases were dried over Na<NUM>SO<NUM>, filtered and 1d was isolated upon evaporation of the solvent. Product 1d was purified using flash chromatography.

To a solution of 1d (<NUM>, <NUM> mmol) in i-PrOH (<NUM>) and H<NUM>O (<NUM>) was added LiOH (<NUM>, <NUM> mmol) and the reaction mixture was stirred for <NUM> at rt. The mixture was diluted with DCM (<NUM>) and washed <NUM> x with <NUM> H<NUM>SO<NUM> (<NUM>) and <NUM> x with brine (<NUM>). The aqueous phases were re-extracted with DCM (<NUM>). The combined organic phases were dried over Na<NUM>SO<NUM>, filtered and 1e was isolated upon evaporation of the solvent. 1e was purified using flash chromatography.

To a solution of 1e (<NUM>, <NUM> mmol) in anhydrous DCM (<NUM>) were added DCC (<NUM>, <NUM> mmol), and a catalytic amount of DMAP. After <NUM>, N-hydroxy-succinimide (<NUM>, <NUM> mmol) was added and the reaction mixture was stirred at rt for <NUM>. The reaction mixture was filtered, the solvent was removed in vacuo and the residue was taken up in <NUM> % acetonitrile plus <NUM> % TFA (<NUM>). The crude mixture was purified by RP-HPLC. Product fractions were neutralized with <NUM> pH <NUM> phosphate buffer and concentrated. The remaining aqueous phase was extracted with DCM and If was isolated upon evaporation of the solvent.

<NUM>-Methoxytriphenylmethyl chloride (<NUM>, <NUM> mmol) was dissolved in DCM (<NUM>) and added dropwise under stirring to a solution of ethylenediamine 2a (<NUM>, <NUM> mmol) in DCM (<NUM>). The reaction mixture was stirred for <NUM> at rt after which it was diluted with diethyl ether (<NUM>), washed <NUM> x with brine/<NUM> NaOH <NUM>/<NUM> (v/v) and once with brine. The organic phase was dried over Na<NUM>SO<NUM> and 2b was isolated upon evaporation of the solvent.

Mmt protected intermediate 2b (<NUM>, <NUM> mmol) was dissolved in DCM (<NUM>). <NUM>-(Tritylthio)-hexanoic acid (<NUM>, <NUM> mmol), PyBOP (<NUM>, <NUM> mmol) and DIPEA (<NUM>, <NUM> mmol) were added and the mixture was stirred for <NUM> at rt. The solution was diuted with diethyl ether (<NUM>), washed <NUM> x with brine/<NUM> NaOH <NUM>/<NUM> (v/v) and once with brine. The organic phase was dried over Na<NUM>SO<NUM> and the solvent was removed in vacuo. 2c was purified using flash chromatography.

Compound 2c (<NUM>, <NUM> mmol) was dissolved in abhydrous THF (<NUM>), <NUM> BH<NUM>·THF solution in THF (<NUM>, <NUM> mmol) was added and the mixture was stirred for <NUM> at rt. More <NUM> BH<NUM>·THF solution in THF (<NUM>, <NUM> mmol) was added and the mixture was stirred for further <NUM> at rt. Methanol (<NUM>) and N,N'-dimethyl-ethylendiamine (<NUM>, <NUM> mmol) were added and the mixture was heated under reflux for <NUM>. The mixture was allowed to cool down and ethyl acetate (<NUM>) was added. The solution was washed <NUM> x with aqueous Na<NUM>CO<NUM> and <NUM> x with aqueous NaHCO<NUM>. The organic phase was dried over Na<NUM>SO<NUM> and the solvent was removed in vacuo to obtain 2d.

Di-tert-butyl dicarbonate (<NUM>, <NUM> mmol) and DIPEA (<NUM>, <NUM> mmol) were dissolved in DCM (<NUM>) and added to a solution of 2d (<NUM>, <NUM> mmol) in DCM (<NUM>). The mixture was stirred for <NUM> at rt. The solution was concentrated in vacuo and purified by flash chromatography to obtain product 2e.

Compound 2e (<NUM>, <NUM> mmol) was dissolved in acetonitrile (<NUM>). <NUM> aqueous HCl (<NUM>) followed by acetonitrile (<NUM>) was added and the mixture was stirred for <NUM> at rt. The pH was adjusted to pH <NUM> by addition of aqueous <NUM> NaOH. The organic solvent was removed in vacuo and the remaining aqueous solution was extracted <NUM> x with DCM. The combined organic phases were dried over Na<NUM>SO<NUM> and the solvent was removed in vacuo to obtain product 2f.

Compound 2f (<NUM>, <NUM> mmol) was dissolved in DCM (<NUM>) and added to a solution of p-nitrophenyl chloroformate (<NUM>, <NUM> mmol) in DCM (<NUM>). <NUM>,<NUM>,<NUM>-trimethylpyridine (<NUM>, <NUM> mmol) was added and the solution was stirred for <NUM> at rt. The solution was concentrated in vacu and purified by flash chromatography to obtain product <NUM>.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus was Fmoc deprotected according to the procedure given in Materials and Methods. A solution of <NUM>-tritylmercaptohexanoic acid (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried in vacuo. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Boc protected N-terminus and ivDde protected side chain of Lys26 was ivDde deprotected according to the procedure given in Materials and Methods. A solution of <NUM>-tritylmercaptohexanoic acid (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried in vacuo. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus was Fmoc deprotected according to the procedure given in Materials and Methods. A solution of Fmoc-Aib-OH (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF and Fmoc-deprotected as described above. A solution of <NUM> (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried in vacuo. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus was Fmoc deprotected according to the procedure given in Materials and Methods. A solution of Fmoc-Ala-OH (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was shaken for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried under vacuum. Fmoc-deprotection was performed as described above. A solution of <NUM> (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried in vacuo. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus was Fmoc deprotected according to the procedure given in Materials and Methods. A solution of Fmoc-Ser(Trt)-OH (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried under vacuum. Fmoc-deprotection was performed as described above. A solution of <NUM> (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to the resin. The suspension was shaken for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried in vacuo. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus was Fmoc deprotected according to the procedure given in Materials and Methods. A solution of Fmoc-Leu-OH (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried under vacuum. Fmoc-deprotection was performed as described above. A solution of <NUM> (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried in vacuo. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Fmoc protected N-terminus was Fmoc deprotected according to the procedure given in Materials and Methods. A solution of 1e (<NUM>, <NUM>µmol), PyBOP (<NUM>, <NUM>µmol) and DIPEA (<NUM>µL, <NUM>µmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried under vacuum. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and agitating the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

Side chain protected PTH(<NUM>-<NUM>) on TCP resin having Boc protected N-terminus and ivDde protected side chain of Lys26 was ivDde deprotected according to the procedure given in Materials and Methods. A solution of 1f (<NUM>, <NUM>µmol) and DIPEA (<NUM>, <NUM> mmol) in DMF (<NUM>) was added to <NUM> (<NUM>µmol) of the resin. The suspension was agitated for <NUM> at rt. The resin was washed <NUM> x with DMF, <NUM> x with DCM and dried under vacuum. Cleavage of the peptide from the resin and removal of protecting groups was achieved by adding <NUM> cleavage cocktail <NUM>/<NUM>/<NUM>/<NUM>/<NUM> (v/w/v/v/v) TFA/DTT/TES/water/thioanisole and shaking the suspension for <NUM> at rt. Crude <NUM> was precipitated in pre-cooled diethyl ether (-<NUM>). The precipitate was dissolved in ACN/water and purified by RP-HPLC. The product fractions were freeze-dried.

<NUM> of a <NUM> NaH<NUM>PO<NUM> buffer (pH <NUM>) was added to <NUM> of a <NUM>/mL solution of thiol <NUM> (<NUM>, <NUM>µmol) in <NUM>/<NUM> (v/v) acetonitrile/water containing <NUM> % TFA (v/v). The solution was incubated at rt for <NUM> after which <NUM>µL of a <NUM>/mL solution of maleimide 11a (<NUM>, <NUM>µmol) in <NUM>/<NUM> (v/v) acetonitrile/water containing <NUM> % TFA (v/v) were added. The solution was incubated for <NUM> at rt. <NUM>µL TFA was added and the mixture was purified by RP-HPLC. The product fractions were freeze-dried to obtain 11b.

Conjugate <NUM> was synthesized as described for 11b by using thiol <NUM> (<NUM>, <NUM>µmol) and maleimide 11a (<NUM>, <NUM>µmol).

<NUM>µL of a solution containing thiol <NUM> (<NUM>/mL, <NUM>, <NUM>µmol) and <NUM>/mL Boc-L-Met in <NUM>/<NUM> (v/v) acetonitrile/water containing <NUM> % TFA (v/v) were added to <NUM> of a solution containing PEG 2x20 kDa maleimide (Sunbright GL2-400MA, <NUM>, <NUM>µmol) and <NUM>/mL Boc-L-Met in water containing <NUM> % TFA (v/v). <NUM> NaH<NUM>PO<NUM> buffer (<NUM>, pH <NUM>) was added and the mixture was stirred for <NUM> at rt. <NUM>µL of a <NUM>/mL solution of <NUM>-mercaptoethanol in water was added. The mixture was stirred for <NUM> at rt and <NUM> <NUM> HCl were added. Conjugate <NUM> was purified by IEX followed by RP-HPLC using a linear gradient of solvent system A (water containing <NUM> % AcOH v/v) and solvent system B (acetonitrile containing <NUM> % AcOH v/v). The product containing fractions were freeze-dried.

Conjugate <NUM> was prepared as described for <NUM> by reaction of thiol <NUM> (<NUM>, <NUM>µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, <NUM>, <NUM>µmol).

Conjugate <NUM> was prepared as described for <NUM> by reaction of thiol <NUM> (<NUM>, <NUM>µmol) and PEG 2x20 kDa maleimide (Sunbright GL2-400MA, <NUM>, <NUM>µmol). The reaction was quenched by addition of <NUM>µL TFA without prior addition of <NUM>-mercaptoethanol. Conjugate <NUM> was purified by IEX followed by SEC for desalting. The product containing fractions were freeze-dried.

Method: This study was conducted in order to test the hypothesis of performing dose adjustments with TransCon PTH in <NUM>% increments in response to hypocalcemia or hypercalcemia. The study was performed in male and female Cynomolgus monkeys. Four groups with one male and one female animal in each were dosed for four weeks starting at doses <NUM>, <NUM>, <NUM> and <NUM>µg/kg/day, respectively. Serum calcium (sCa) levels in the animals were measured pre-dose and post-dose on days -<NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM> and <NUM>.

Results: Daily subcutaneous administration of TransCon PTH at doses of <NUM>, <NUM> and <NUM>µg/kg/day for <NUM> days was well tolerated and was associated with expected sporadic slight increases in sCa levels (from <NUM>. 05x up to <NUM>. 12x compared to pre-study levels). On Day <NUM> for the male and female dosed at <NUM>µg/kg/day day sCa rose to severe hypercalcaemic levels (up to <NUM>. 42x and <NUM>. 34x, respectively, compared to pre-study levels). Dosing was discontinued in these two animals followed by a dosing holiday. On Day <NUM> dosing was resumed at <NUM>µg/kg/day (<NUM>% dose reduction) for one week. On Day <NUM> (first day of dosing after the dosing holiday), increases in calcium were noted at all timepoints for the male (from <NUM> x to <NUM> x to pre-study levels) with peak concentration achieved <NUM> hrs postdose. On Day <NUM>, the increases were noted for male and female (up to <NUM> x to pre-study levels). On Day <NUM>, calcium increases were still present in both sexes.

Claim 1:
A pharmaceutical composition comprising a controlled-release PTH compound or a pharmaceutically acceptable salt, hydrate, or solvate thereof, for use in a method of treating hypoparathyroidism, wherein the method follows a dosage regimen in which dose adjustment in response to hypocalcemia or hypercalcemia is performed in increments of no more than <NUM>%, and wherein the controlled-release PTH compound releases at least one PTH molecule with a release half-life of at least <NUM> hours in aqueous buffer, pH <NUM> and <NUM>, wherein the controlled-release PTH compound is a compound of formula (Ia)
<CHM>
wherein -Z comprises a moiety of formula (b)
<CHM>
wherein
the dashed line indicates attachment to -L<NUM>- or to the remainder of -Z; and
m and p are independently of each other an integer ranging from and including <NUM> to <NUM>, and
-L<NUM>-L<NUM>- has the formula:
<CHM>
wherein the unmarked dashed line indicates attachment by an amide bond to a nitrogen of -D, and the dashed line marked with an asterisk indicates attachment to -Z, wherein -D has the sequence of SEQ ID NO:<NUM> and x is <NUM>.