WO2012120532A2 - Peptides cycliques à chaîne courte - Google Patents

Peptides cycliques à chaîne courte Download PDF

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WO2012120532A2
WO2012120532A2 PCT/IN2012/000078 IN2012000078W WO2012120532A2 WO 2012120532 A2 WO2012120532 A2 WO 2012120532A2 IN 2012000078 W IN2012000078 W IN 2012000078W WO 2012120532 A2 WO2012120532 A2 WO 2012120532A2
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aib
phe
ame
har
nle
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PCT/IN2012/000078
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WO2012120532A3 (fr
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Rajesh H Bahekar
Vjay PRAJAPATI
Mukul R. Jain
Pankaj R. Patel
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Cadila Healthcare Limited
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/635—Parathyroid hormone (parathormone); Parathyroid hormone-related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to novel cyclic short chain peptides of general formula (I) having beneficial pharmacological properties, their pharmaceutically acceptable salts and pharmaceutical compositions containing them.
  • the present invention also relates to processes for preparing compounds of general formula (I), their pharmaceutically acceptable salts and pharmaceutical compositions containing them.
  • Osteoporosis is a skeletal disorder characterised by diminished bone mass, decreased bone mineral density (BMD), decreased bone strength and associated with an increased risk of bone fracture (Lane J. M., et al., Clin. Orthop. Relat. Res., 372, 2000, 139-150). Osteoporotic fractures most often occur in the vertebrae, hips or the femoral neck. These fractures severely impair the quality of life because of pain, long-lasting immobility and poor recovery. Bone comprises of several different cell types. Osteoblast (bone formation) lays down new bone from the mineral present in the extracellular milieu around the cells. Osteoclasts (bone loss) remove old bones, releasing the minerals compiled within bone back into the extracellular matrix.
  • BMD bone mineral density
  • Osteoporosis occurs when the rate of the bone resorption is greater than the rate of bone formation (Seeman E., et al., N. Engl. J. Med., 354(21), 2006, 2250-2261).
  • Postmenopausal estrogen deficiency is the most common cause of osteoporosis in women, as estrogen puts a break on osteoclast lifespan.
  • Other major risk factors in the development of osteoporosis include: low calcium intake, vitamin D deficiency, type-1 diabetes, rheumatoid arthritis, long-term use of medication such as anticonvulsants and corticosteroids and low levels of testosterone in men (Cole Z.
  • PTH Native human Parathyroid Hormone
  • PTH in-turn, maintains the serum calcium level by directly or indirectly promoting calcium entry into the blood.
  • PTH contributes to net gastrointestinal absorption of calcium by favouring the renal synthesis of the active form of vitamin D.
  • PTH promotes calcium reabsorption from bone, indirectly by stimulating differentiation of the osteoclasts (bone-resorbing cells).
  • Administration of PTH via parenteral route efficiently increases bone mineral density (BMD), bone strength and reduces the incidence of new osteoporotic fractures in osteoporotic patients (Greenspan S. L., et al., Ann. Intern. Med., 146(5), 2007, 326- 339; Neer R. M., et al., N. Engl. J. Med., 344, 2001, 1434-1441).
  • PTH receptor which is expressed in numerous tissues, most abundantly in kidney, bone and growth plate chondrocytes (Lanske B., et al., Crit. Rev. Eukaryot. Gene Expr., 8, 1998, 297-320).
  • the PTH receptor is homologous in primary structure to a number of other receptors that bind peptide hormones, such as secretin, calcitonin and glucagon; together, these receptors form a distinct family called G-protein coupled receptors (GPCR/ GPCRs) family B (Kolakowski L. F., Receptor Channels, 2, 1994, 1-7).
  • the GPCR comprise an extracellular N-terminal domain of 100-160 residues, connected to a juxtamembrane domain (J-domain) of seven membrane-spanning a-helices with intervening loops and a C-terminal tail (Donnelly D., FEBS Letts., 409, 1997, 431- 436).
  • the Class B GPCRs are activated by endogenous peptide ligands of intermediate size, typically 30-40 amino acids (Hoare, S. R. J., Drug Discovery Today, 10, 2005, 423-427). A general mechanism of peptide ligand interaction with class B GPCRs has emerged and is termed as the 'two-domain' model.
  • the C-terminal portion of the peptide binds the N-domain of the receptor, confirm binding of ligand with the receptor and the N-terminal ligand region binds the J-domain, an interaction that activates the receptor and stimulates intracellular signalling (Ji T. H., et al., J. Biol. Chem., 273, 1988, 17299-17302; Hjorth, S. A., et al., Regulatory Peptides, 64, 1996, 70).
  • PTH binds to the PTH receptor with affinity in the nM range; the ligand- occupied receptor transmits a signal across the cell membrane to intracellular effector enzymes through a mechanism that involves intermediary heterotrimeric GTP-binding proteins (G proteins).
  • G proteins GTP-binding proteins
  • the primary intracellular effector enzyme activated by the PTH receptor in response to PTH peptide is adenylyl cyclase (AC) (Goltzman D., J. Bone Miner. Res., 15(3), 2000, 605-608).
  • PTH induces increase in the second messenger, cyclic adenosine monophosphate (cAMP) which regulates the poorly characterized downstream cellular processes involved in bone remolding (Juppner H., et al., Science, 254, 1991, 1024-1026).
  • cAMP cyclic adenosine monophosphate
  • Other signalling pathways of this receptor such as elevation of intracellular calcium, phospholipase C-dependent and independent activation of protein kinase C, have been described.
  • the parathyroid hormone and its derivatives represent potential therapeutic agent for the treatment of osteoporosis (Slovik D. M., et al., J. Bone Miner. Res., 1, 1986, 377-381 ; Dempster D. W., et al., Endocr. Rev., 14, 1993, 690-709).
  • Synthetic PTH (1 -34) exhibits full bioactivity in most cell-based assay systems, has potent anabolic effects on bone mass in animals and has recently been shown to reduce the risk of bone fracture in postmenopausal osteoporotic women.
  • daily subcutaneous injections of low doses of PTH (1-34) were shown to result in impressive bone formation in the spine and femoral neck with significant reduction in incidence of vertebral fractures (Neer R. M., et al., N. Engl. J. Med., 344, 2001, 1434-1441; Dempster D. W., et al., Endocr. Rev., 14, 1993, 690-709).
  • PTH as one of the most efficacious agents tested for osteoporosis.
  • PTH (1-34) in the form of teriparatide acetate has been approved for the treatment of osteoporosis.
  • PTH derivatives include polypeptides that have amino acid substitutions or are truncated relative to the full-length molecule. Both the N and C-terminal truncated forms of PTH (1-34) has been studied. Additionally, amino acid substitutions within the truncated polypeptides have also been investigated. (Azurani A., et al., J. Biol. Chem., 271, 1996, 14931-14936).
  • Truncated PTH (1-34) derivatives such as cyclised PTH (1-17), PTH (1-28), PTH (1 -31) and PTH (1 -1 1 ) are active in most assay systems and promote bone-formation (Whitfield J. F., et al., J. Bone Miner. Res., 12, 1997, 1246-1252; WO 2007 / 1301 13 A2; WO 2008 / 068487; Whitefild J. F., et al., Calcif. Tissue Int., 56, 1995, 227-231 ; Rixon R. H., et al., J. Bone Miner.
  • PTH is highly susceptible to protease attack and must be stored at low temperature due to its low stability.
  • tolerability is limited by transient mobilization of calcium and hypercalcemia also the toxicological data and in particular the unfavourable results of cancerogenesis studies (dose and treatment duration dependent increased risk of osteosarcoma) induce a cautious use of PTH (1 - 34) (Vahle J. L., Toxicol. Pathos., 32(4), 2004, 426-438; Whitfiel J.
  • PTH(l-l l) analogs [Ala 3 , Glnio, Argn]-PTH(1-1 1), [Ala 3 , Glnio, Har, ,]-PTH(l -l l) and [Aib 1 >3 ; Gln, 0 ; Harness]-PTH(1-1 1)
  • PTH (1-14) analogs such as [AC5C1, Aib 3 , Glnio, Harn, Ala )2 , Trpi ]PTH(l-14) stimulate cAMP, in nM range (WO 03/009804; WO 04/093902).
  • short chain peptide analogues consisting of 1 1 amino acids (derivatives of first 1-1 1 residues of PTH peptide, Seq. ID. No.2) can activate the PTH receptor (in vitro) with low potency (WO 04/067021 ), however, in animal models (in vivo) bone-anabolic activity of these analogues has not been reported.
  • agonistic activity on cAMP- signalling pathway of the PTH receptor (in vitro) alone is not at all predictive for bone- anabolic activity in vivo.
  • PTH (1-15) 'SVSEIQLMHNLGKHL 15 (Seq. ID No: 3)
  • PTH parathyroid hormone peptide
  • Xaal and Xaa3 represent either Aib or AC 5 C
  • Xaa8 represent Nle
  • XaalO represent S & E
  • Xaal l represent Har
  • Xaal 2 represent Ala
  • Xaal4 represent W (WO 03 / 009804 A2; US 2006 / 7153951 B2; US 2007 / 01 17157 Al ; US 2007 / 0203071 Al ; US 2006 / 0019902 Al ; US 2007 / 0 . 161569 Al ; US 2007 / 011 1946 Al ; Gardella T. J., et al., J. Biol. Chem. 2000, 275, 21836-21843; Gardella T.
  • the present invention describes a group of novel cyclic short chain peptides that function as agonist of the PTH receptor, having different degree of affinity towards the PTH / PTH-1 receptor and useful for the treatment of osteoporosis.
  • These cyclic peptides are defined by the general formula (I) as given below.
  • the cyclic peptides of the present invention are useful for the treatment or prevention of hypoparathyroidism and diseases characterized by bone mass reduction or bone loss, such as osteoporosis, postmenopausal osteoporosis and for stimulating bone repair.
  • the present invention provides novel cyclic peptides of formula (I), which primarily act as a PTH/ PTH-1 receptor agonist. These cyclic peptides exhibit increased metabolic stability against proteolytic enzymes. Most of these cyclic peptides were found to be stable in rat plasma upto 24 hours (in vitro), showed increased stability against GIT enzymes such as pepsin and acidic stomach pH and also against liver microsomes (in vitro). Due to increased metabolic stability, other than parenteral route of administration, most of these cyclic peptides can also be delivered by oral route of administration.
  • a preferred embodiment of the present invention is to provide novel cyclic peptides of general formula (I), novel intermediates involved in their synthesis, their pharmaceutically acceptable salts and pharmaceutical compositions containing them or their mixtures, suitable for the treatment/mitigation/regulation of bone disorders.
  • compositions containing cyclic peptides of general formula (I), their pharmaceutically acceptable salts, solvates and their mixtures having pharmaceutically acceptable carriers, solvents, diluents, excipients and other components normally employed in their manufacture are provided.
  • novel cyclic peptides of the present invention as agents for stimulating new bone formation and treating and / or preventing osteoporosis and related bone disorders, by administering a therapeutically effective and non-toxic amount of the cyclic peptides of formula (I), or their pharmaceutically acceptable compositions to the mammals those are in need of such treatment.
  • Aib a-Amino-isobutyric acid
  • aMe-APPA 2-amino-2-methyl-5-phenylpentanoic acid
  • ccMe-Bip(OMe) alpha-methylated Bip(OMe)
  • aMe-Pro alpha-methyl-Proline
  • aMe-Phe alpha-methyl-phenylalanine
  • aMe-2F-Phe alpha-methyl-2-fluorophenylalanine
  • a e-2,6-F-Phe alpha-methyl-2,6-diflurophenylalanine
  • ccMe-Trp alpha-methyl-Tryptophan
  • aMe-Tyr alpha-methyl-Tyrosine
  • ACN Acetonitrile
  • cAMP Adenosine 3',5'-cyclic monophosphate
  • DIPCDI Di-isopropylcarbodiimide
  • DIPEA Diisopropylethylamine
  • Fmoc Fluorenylmethoxycarbonyl
  • HoSer Homoserine
  • HOBt 1-Hydroxybenzotriazole
  • HOAt 7-Aza-hydroxybenzotriazole
  • HBTU 2-(lH-benzotriazole-l-yl)-l,l,3,3-tetramethyl aminium hexafluorophosphate
  • HPLC High Performance Liquid Chromatography
  • PTH Parathyroid Hormone
  • PTH-lr agonist Parathyroid Hormone receptor agonist
  • PyBOP Benzotriazole-l -yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate
  • Pyr Pyroglutamic acid
  • TIPS Triisopropylsilane
  • TFA Trifluoroacetic acid
  • Figure 1 Examples of orthogonally protected amino acids used in Fmoc based-solid phase peptide synthesis (SPPS) of short-chain peptides.
  • SPPS Fmoc based-solid phase peptide synthesis
  • Figure 2 In vitro DRC and EC 50 determination of PTH(l-34) ( Figure A) and Seq. ID No. 133 ( Figure B), in Rat PTH-1 R assay (agonistic activity, measured by amount of cAMP released).
  • Figure 3 In vivo DRC study with Seq. ID No. 168 (Figure A: % increase in serum Ca 2+ levels; Figure B: % decrease in serum P0 4 levels), in OVX Female Rat.
  • Figure 4 Changes in biochemical parameters and femur weight, in OVX Rat after 6 weeks treatment with Seq. ID No. 247 and PTH(l-34).
  • synthetic cyclic short chain peptides having the structural formula (I), which showed PTH-1 receptor agonistic activity are disclosed. These cyclic short chain peptides exhibit increased metabolic stability to proteolytic cleavage, as most of cyclic peptides were found to be stable in rat plasma upto 24 hours (in vitro), showed increased stability against GIT enzymes such as pepsin and acidic stomach pH and also against liver microsomes (in vitro).
  • cyclic peptides can also be delivered by oral routes of administration, for the treatment / prevention of hypoparathyroidism and diseases characterized by bone mass reduction, such as osteoporosis, postmenopausal osteoporosis and for stimulating bone repair.
  • the present invention thus discloses novel cyclic short chain peptides as PTH receptor agonist having the following structure (I).
  • ⁇ ' represents the groups -NH-Ri or R3-CO-NH- , wherein Ri represents hydrogen or optionally substituted linear (Cns) alkyl chain, or suitable amino acids preferably selected from pyroglutamic acid (Pyr), Pro (P), alpha-methyl-Proline (aMe-P), Val (V), alpha-methyl-Valine (aMe-V); R 3 is selected from optionally substituted linear (Ci-] 8 ) alkyl chain, (Cr 6 )alkoxy, (C 3 -C 6 ) cycloalkyl, aryl, heteroaryl or arylalkyl groups;
  • the aryl group is selected from phenyl, napthyl, indanyl or biphenyl, groups; the heteroaryl group is selected from pyridyl, thienyl, furyl, imidazolyl, benzofuranyl groups; the arylalkyl groups represent groups wherein the aryl group is attached to an alkyl groups as defined elsewhere in the specification.
  • 'B' represents -COOR 2 , -CONHR 2 or CH 2 OR 2, wherein R 2 represents H or suitable amino acids.
  • Preferred amino acids may be selected from the group comprising of Val (V), alpha-methyl-Valine (aMe-V), and the like;
  • Each of Zi ; Z 3 & Z 2 may be same or different and independently represents a naturally or unnaturally occurring amino acids selected from the group comprising of Ser (S), Val (V), Pro (P), alpha-methyl-Proline (aMe-P), Gly (G), Ala (A), a-amino- isobutyric acid (Aib), 1 -amino cyclopropane carboxylic acid (AC 3 C), 1-amino- cyclopentanecarboxylic acid (AC 5 C), 1-amino-cyclohexanecarboxylic acid (AC 6 C); preferably, each of Zi, Z 3 & Z ) represents amino acid selected from Ala (A), a-amino- isobutyric acid (Aib), 1-amino cyclopropane carboxylic acid (AC 3 C) or 1-amino- cyclopentanecarboxylic acid (AC5C);
  • Z 2 represents either a Val (V) or alpha-methyl-valine (aMe-V);
  • Z4 represents a naturally or unnaturally occurring amino acids selected from the group comprising of Glu(E), Asp(D).
  • Each of Z 6 & Z10 may be same or different and independently represents a naturally or unnaturally occurring amino acid selected from the group comprising of Ser (S), Homoserine (HoSer), Lys (K), Orn (Ornithine), Glu (E), Homoglutamic acid (HoGlu), Asp (D); wherein, the side chains of Z 6 and Z10 having acidic or basic group such as Lys (K), Glu (E) or Asp (D) lead to the formation of lactam bridge or the side chain of Z 6 and Z ]0 having alcohol group such as Ser (S) or Homoserine (HoSer) are cross-linked as a diester bridge, with dicarboxylic acids, such as succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid or sebacic acid; preferably each of Z 6 & Z10 represents amino acid selected from Ser (S), Lys (K), Glu (E) or Asp (D);
  • Z 8 represents a naturally or unnaturally occurring amino acid selected from the group comprising of Met, alpha-methyl-Valine (ocMe-V), Leu, Nle, HoLeu, Val; preferably Z 8 represents amino acid selected from Met, Leu or Val;
  • ] 1 & Zi 3 may be same or different and independently represents a naturally or unnaturally occurring amino acid selected from the group comprising of Lysine (K), Arginine (R), Homoarginine (Har), Ornithine (Orn), Citrulline (Cit), Phenyl alanine (F), alpha-methyl-phenylalanine (aMe-Phe-), alpha-methyl-2-fluorophenylalanine (ocMe-2F-Phe-) or alpha-methyl-2,6-difIurophenylalanine (o Me-2,6-F-Phe-) or 2- fluorophenylalanine (2F-Phe-); preferably, each of Zn & Zj 3 represents amino acid selected from Lysine (K), Arginine (R), Homoarginine (Har) or Citrulline (Cit);
  • Zn may be present or absent.
  • Zj 4 when present represents a naturally or unnaturally occurring amino acid selected from the group comprising of 2'-ethyl-4'- meihoxy-biphenylalanine (Bip(OMe)), ot-methylated Bip(OMe) [aMe-Bip(OMe)], alpha-methyl-tryptophan (ocMe-Trp), alpha-methyl-tyrosine (aMe-Tyr), Tryptophan (W), Tyrosine (Y), D-Tryptophan ( ⁇ /W), D-Tyrosine (dY), alpha-methyl-phenylalanine (aMe-Phe-), alpha-methyl-2-fluorophenylalanine (aMe-2F-Phe-), alpha-methyl-2,6- diflurophenylalanine (aMe-2,6-F-Phe-) or 2-fluorophenylalanine (2F-Phe-); preferably
  • Zi5 may be present or absent.
  • Zj 5 when present represents a naturally or unnaturally occurring amino acid selected from the group comprising of 2-aminb-5- phenylpentanoic acid (APPA) or 2-amino-2-methyl-5-phenylpentanoic acid (aMe- APPA);
  • APPA 2-aminb-5- phenylpentanoic acid
  • aMe- APPA 2-amino-2-methyl-5-phenylpentanoic acid
  • 'unnatural amino acids' or 'non-natural amino acids' preferably represents either replacement of L-amino acids with corresponding D-amino acids such as replacement of L-Ala with D-Ala and the like or suitable modifications of the L or D amino acids, amino alkyl acids, either by
  • - a-alkylation such as substitution of Ala with cc-methyl Ala (Aib), replacement of Met with -methyl Met;
  • - substitution on the side chain of amino acid such as substitution of aromatic amino acid side chain with halogen, (C1-C3) alkyl, aryl groups, more specifically the replacement of Phe with halo Phe;
  • alkyl used herein, either alone or in combination with other radicals, denotes a linear or branched radical containing one to eighteen carbons, such as methyl, ethyl, H-propyl, w-butyl, amyl, «-pentyl, w-hexyl, heptyl, octyl, decyl, tetradecyl, octadecyl and the like.
  • cycloalkyl used herein, either alone or in combination with other radicals, denotes a radical containing three to seven carbons, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
  • 'amino acid' as employed herein alone or as part of another group includes, without limitation, an amino group and a carboxyl group linked to the same carbon, referred to as 'a' carbon.
  • the absolute 'S' configuration at the 'a' carbon is commonly referred to as the 'L' or natural configuration.
  • the 'R' configuration at the 'a' carbon is commonly referred to as the 'D' amino acid.
  • both the 'a-substituents' is equal, such as hydrogen or methyl, the amino acids are Gly or Aib and are not chiral.
  • the compounds of the invention include peptide amides and non-amides and peptide analogues, including but not limited to the following:
  • 'PTH receptor modulator or agonist' refers to a compound that acts at the PTH-1 and/or PTH-2 receptor to alter its ability to regulate downstream signaling events, such as cAMP production.
  • Example of receptor modulators includes agonist, partial agonist, inverse agonist and allosteric potentiators.
  • the synthetic isolated cyclic peptides described herein primarily acts as PTH receptor agonists.
  • These synthetic cyclic peptides exhibit desirable in vitro PTH receptor agonist activity in UMR-106 cells, in the range of 1-1000 nM concentration. PTH receptor agonistic activity is assessed by estimation of amount of cAMP released by the test compounds.
  • Some of the cyclic peptides prepared showed increase in BMD and / or bone strength, when tested in vivo, in OVX rat model, thus making them ideal therapeutic candidates for the treatment and prevention of osteoporosis.
  • These new classes of cyclic peptides can be administered by oral or other non-invasive routes or parenteral routes of administration.
  • the present invention provides cyclic short chain peptides of formula (I) pharmaceutical compositions employing such cyclic short chain peptides either alone or in combination and for methods of using such cyclic peptides.
  • the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of cyclic peptides of formula (I), alone or in combination(s), with a pharmaceutically acceptable carrier.
  • a method for treating or delaying the progression or onset of osteoporosis especially primary osteoporosis, endocrine osteoporosis, postmenopausal osteoporosis, hereditary and congentinal forms of osteoporosis, wherein, therapeutically effective amount of cyclic peptides of formula (I) or their combination(s) can be administered to a mammal, example, human and a patient in need of treatment.
  • cyclic short chain peptides of the present invention can be synthesized using the methods described below, together with conventional techniques known to those skilled in the art of peptide synthesis, or variations thereon as appreciated by those skilled in the art. Referred methods include, but not limited to those described below.
  • the cyclic short chain peptides thereof described herein may be produced by chemical synthesis using suitable variations of both the solution-phase (preferably, using Boc- chemistry; as generally described in M. Bodansky, A. Bodansky, "The practice of peptide synthesis", Springer-Verlag, Berlim, 1984; E. Gross, J. Meinhofer, "The peptide synthesis, analysis, biology", Vol. 1, Academic Press, London, 1979) and or solid-phase techniques, such as those described generally in G. Barany & R. B. Merrifield, "The peptides: Analysis, synthesis, Biology”; Volume 2- "Special methods in peptide synthesis, Part A", pp. 3-284, E. Gross & J. Meienhofer, Eds., Academic Press, New York, 1980; and in J. M. Stewart and J. D. Young, "Solid-phase peptide synthesis” 2nd Ed., Pierce chemical Co., Rockford, II, 1984.
  • the preferred strategy for preparing the cyclic peptides of this invention is based on the use of Fmoc-based SPPS approach, wherein Fmoc (9- fluorenylmethoxycarbonyl) group is used for temporary protection of the a-amino group, in combination with the acid labile protecting groups, such as tert- butoxycarbonyl (Boc), tert-butyl (Bu l ), Trityl (Trt) groups ( Figure 1), for temporary protection of the amino acid side chains, if present (see for example E. Atherton & R.C.
  • the cyclic short chain peptides can be synthesized in a stepwise manner on an insoluble polymer support (resin), starting from the C-terminus of the peptide.
  • the synthesis is initiated by appending the C-terminal amino acid of the peptide to the resin through formation of an amide, ester or ether linkage. This allows the eventual release of the resulting peptide as a C-terminal amide, carboxylic acid or alcohol, respectively.
  • the C-terminal amino acid and all other amino acids used in the synthesis are required to have their a-amino groups and side chain functionalities (if present) differentially protected (orthogonal protection), such that the a-amino protecting group may be selectively removed during the synthesis, using suitable base such as 20% piperidine solution, without any premature cleavage of peptide from resin or deprotection of side chain protecting groups, usually protected with the acid labile protecting groups.
  • the coupling of an amino acid is performed by activation of its carboxyl group as an active ester and reaction thereof with unblocked a-amino group of the N-terminal amino acid appended to the resin. After every coupling and deprotection, peptidyl-resin was washed with the excess of solvents, such as DMF, DCM and diethyl ether. The sequence of a-amino group deprotection and coupling is repeated until the desired peptide sequence is assembled (Scheme 1). The peptide is then cleaved from the resin with concomitant deprotection of the side chain functionalities, using an appropriate cleavage mixture, usually in the presence of appropriate scavengers to limit side reactions. The resulting peptide is finally purified by reverse phase HPLC.
  • the synthesis of the peptidyl-resins required as precursors to the final peptides utilizes commercially available cross-linked polystyrene polymer resins (Novabiochem, San Diego, CA).
  • Preferred for use in this invention is Fmoc-PAL-PEG-PS resin, 4-(2', 4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetyl-j?-methyl benzhydrylamine resin (Fmoc-Rink amide MBHA resin), 2-chloro-Trityl-chloride resin or p- benzyloxybenzyl alcohol resin (HMP resin) to which the C-terminal amino acid may or may not be already attached.
  • Fmoc-PAL-PEG-PS resin 4-(2', 4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetyl-j?-methyl benzhydrylamine resin
  • the C-terminal amino acid is not attached, its attachment may be achieved by HOBt active ester of the Fmoc-protected amino acid formed by its reaction with DIPCDI.
  • HOBt active ester of the Fmoc-protected amino acid formed by its reaction with DIPCDI.
  • 2-Chloro-trityl resin coupling of first Fmoc-protected amino acid was achieved, using DIPEA.
  • N- terminal protection of peptidyl resin was selectively deprotected using 10-20 % piperidine solution. After every coupling and deprotection, excess of amino acids and coupling reagents were removed by washing with DMF, DCM and ether.
  • Coupling of the subsequent amino acids can be accomplished using HOBt or HOAt active esters produced from DIPCDI/ HOBt or DIPCDI/HOAt, respectively.
  • HOBt or HOAt active esters produced from DIPCDI/ HOBt or DIPCDI/HOAt, respectively.
  • complete coupling can be achieved using a combination of highly efficient coupling agents such as HBTU, PyBOP or TBTU, with additives such as DIPEA.
  • the synthesis of the cyclic peptides described herein can be carried out by using batchwise or continuous flow peptide synthesis apparatus, such as CS-Bio or AAPPTEC peptide synthesizer, utilizing the Fmoc/trityl protection strategy.
  • the non- natural non-commercial amino acids present at different position were incorporated into the peptide chain, using one or more methods known in the art.
  • Fmoc- protected non-natural amino acid was prepared in solution, using appropriate literature procedures.
  • Fmoc-protected APPA analogs were prepared from L-pyroglutamic acid, in good enantiomeric purity, using modified literature procedure (Betsbrugge J.V., et al., Tetrahedron, 54, 1988, 1753- 1762).
  • the Fmoc-protected cc-methylated amino acids were prepared using asymmetric Strecker synthesis (Boesten, W.H.J., et al., Org. Lett., 3(8), 2001, 1 121 -1 124; Cativiela C, Diaz-de-villegas M. D., Tetrahedran Asymmetry, 9, 1988, 3517-3599).
  • the resulting derivative was then used in the step-wise synthesis of the peptide.
  • the required non-natural amino acid was built on the resin directly using synthetic organic chemistry procedures and a linear peptide chain were building.
  • Figure 1 Examples of some of the protected amino acids used in Fmoc based-solid phase peptide synthesis (SPPS) of short-chain peptides.
  • SPPS Fmoc based-solid phase peptide synthesis
  • the peptide-resin precursors for their respective cyclic peptides may be cleaved and deprotected using suitable variations of any of the standard cleavage procedures described in the literature (King D.S., et al., Int. J. Peptide Protein Res., 1990, 36, 255- 266).
  • a preferred method for use in this invention is the use of TFA cleavage mixture, in the presence of water and TIPS as scavengers.
  • the peptidyl-resin was incubated in TFA / Water /TIPS (95:2.5:2.5) for 1.5 - 4 hrs at room temperature.
  • the cleaved resin is then filtered off and the TFA solution is concentrated or dried under reduced pressure.
  • the resulting crude peptide is either precipitated or washed with Et 2 0 or is re-dissolved directly into DMF or 50 % aqueous acetic acid for purification by preparative HPLC.
  • the cyclic short chain peptides with the desired purity can be obtained by purification using preparative HPLC.
  • the solution of crude peptide is injected into a semi-Prep column (Luna 10 ⁇ ; C] 8 ; 100 A ), dimension 250 X 50 mm and eiuted with a linear gradient of ACN in water, both buffered with 0.1 % TFA, using a flow rate of 40 mL /min with effluent monitoring by PDA detector at 220 nm.
  • the structures of the purified cyclic peptides can be confirmed by Electrospray Mass Spectroscopy (ES-MS) analysis.
  • ES-MS Electrospray Mass Spectroscopy
  • peptide prepared were isolated as trifluoro-acetate salt, with TFA as a counter ion, after the Prep-HPLC purification.
  • some peptides were subjected for desalting, by passing through a suitable ion exchange resin bed, preferably through anion-exchange resin Dowex SBR P(C1) or an equivalent basic anion-exchange resin.
  • TFA counter ions were replaced with acetate ions, by passing through suitable ion-exchange resin, eiuted with dilute acetic acid buffer.
  • selected peptides, with the acetate salt was treated with 4 M HC1.
  • the Fmoc-protection on first amino acid, coupled with resin was deprotected, using a 10-20% piperidine solution, followed by the coupling the Fmoc-protected second amino acid, using a suitable coupling agents, and as described above.
  • the repeated cycles of deprotection, washing, coupling and washing were performed until the desired peptide chain was assembled on resin, as per general Scheme 1 above.
  • Aib-V-Aib-EI -(aMe-2F-Phe)-Nle-HE-Har-A-Cit-( Me-2F-Phe) was assembled on an automated CS-Bio 536 PepSynthesiserTM using Fmoc solid phase peptide synthesis (SPPS) approach (Scheme 2).
  • SPPS Fmoc solid phase peptide synthesis
  • the Fmoc amino acids and the 2-(lH-Benzotriazol-l- yl)-l,l,3,3-tetramethyluroniumtetrafluoroborate (TBTU) were packed together in vials and positioned in the amino acid module of the synthesizer.
  • a stock solution of diisopropylethylamine (DIPEA; 0.9 M) and DMF were stored in reagent bottles, under dry nitrogen atmosphere.
  • the resin, Fmoc-PAL-PEG- PS (0.38 mmol/g; lg) was dried over P 2 0 5j in vacuo (I hr) and swollen in freshly distilled DMF (5 mL).
  • the swollen resin was slurry packed into a glass column and positioned in the synthesizer. All the synthetic cycles were carried out at a flow rate of 5 mL min "1 , Table 1.
  • the resin was washed with freshly distilled DMF for 10 minutes.
  • Deprotection of Fmoc group was performed with 20% piperidine in DMF for 10 minutes and the deprotection was monitored by UV detection of the column effluent at 304 nm.
  • the diester bridge formation was carried out using dicarboxylic acid (1-2 eq.), HBTU (4-6 eq.), HOAt (4-6 eq.), and collidine (6-8 eq.), while lactam bridge was prepared by treating the peptide resin, with solution of HBTU (4-6 eq.), HOBt (4-6 eq.), and DIPEA (8-10 eq.).
  • the dried Peptide-resin (ca 10-mg) was treated with mixture (1 mL) of TFA, water, triisopropylsilane (95: 2.5: 2.5 v/v), for 90 minutes at room temperature with gentle occasional swirling.
  • the resin was filtered, washed thoroughly with neat TFA (1 mL) and the entire filtrate was evaporated under reduced pressure. Residual TFA was azeotroped three times with diethyl ether (2 mL). Residue obtained was suspended in distilled water (2 mL) and the aqueous layer was extracted three times with diethyl ether (3 mL). The aqueous layer was separated and freeze-dried to yield the crude
  • the lyophilised peptide Aib-V-Aib-EIK-(aMe-2F-Phe)-Nle-HE-Har-A-Cit-(otMe-2F-Phe) was dissolved in 0.1% aqueous TFA (ca lmg /l mL) and its purity was analyzed by analytical RP-HPLC and characterized by electrospray ionisation mass spectrometry (ESI-MS). Percent purity: 90 % (crude peptide). ESI-MS; Calcd.
  • cyclic short-chain peptides designed in the present invention were prepared, using Fmoc-SPPS approach. Furthermore, resin bound cyclic short-chain peptides were cleaved and deprotected, purified and characterized using following protocol.
  • the desired cyclic short-chain peptides were cleaved and deprotected from their respective peptidyl-resins by treatment with TFA cleavage mixture as follows.
  • a solution of TFA / Water / Triisopropylsilane (95: 2.5: 2.5) (10 mL / 100 mg of peptidyl- resin) was added to peptidyl-resins and the mixture was kept at room temperature with occasional starring.
  • the resin was filtered, washed with a cleavage mixture and the combined filtrate was evaporated to dryness.
  • a solution of crude peptide dissolved in DMF or water was injected into a semi-Prep column (Luna 10 ⁇ ; Ci 8 ; 100 A 0 ), dimension 250 X 50 mm and eluted with a linear gradient of ACN in water, both buffered with 0.1 % TFA, using a flow rate of 15 -50 mL / min, with effluent monitoring by PDA detector at 220 nm.
  • a typical gradient of 20 % to 70 % of water-ACN mixture, buffered with 0.1 % TFA was used, over a period of 50 minutes, with 1% gradient change per minute.
  • each peptide was analyzed by analytical RP-HPLC on a Shimadzu LC-10AD analytical HPLC system.
  • analytical HPLC analysis of cyclic short-chain peptides Luna 5 ⁇ ; Ci 8 ; 100 A ° , dimension 250 X 4.6 mm column was used, with a linear gradient of 0.1% TFA and ACN buffer and the acquisition of chromatogram was carried out at 220 nm, using a PDA detector. .
  • Each peptide was characterized by electrospray ionisation mass spectrometry (ESI-MS), either in flow injection or LC/MS mode.
  • ESI-MS electrospray ionisation mass spectrometry
  • Triple quadrupole mass spectrometers API-3000 (MDS-SCIES, Canada) was used in all analyses in positive and negative ion electrospray mode. Full scan data was acquired over the mass range of quadrupole, operated at unit resolution. In all cases, the experimentally measured molecular weight was within 0.5 Daltons of the calculated monoisotopic molecular weight. Quantification of the mass chromatogram was done using Analyst 1.4.1 software.
  • novel cyclic short chain peptides were prepared. This list is indicative of the various groups of cyclic short chain peptides, which can be prepared according to the present invention, and are expected to at least include obvious variations of these cyclic short chain peptides. However, such disclosure should not be construed as limiting the scope of the invention in any way.
  • Table 2 (i-xvi) novel cyclic short chain peptides of present invention are listed along with their corresponding Seq. ID. No.
  • Table 2 (vi): List of cyclic short chain peptides prepared including mass & % purity for several of the peptides prepared.
  • liver microsomes (ex-vivo).
  • the PTHR is a GPCR and the PTHR agonist binds to it and through signal transduction causes activation of adenylate cyclase and raises intracellular cAMP levels.
  • UMR-106 rat osteosarcoma cells Source ATCC
  • UMR-106 cells were cultured in DMEM nutrition media (Sigma) containing
  • FBS Fetal Bovine Serum
  • IX Penstrep 10% Fetal Bovine Serum (FBS) and IX Penstrep.
  • Cultured cells were plated at a density of 5 x 10 4 cells per well in DMEM media supplemented with 10% FBS, in 96- well tissue culture plates and grown for 2 days before the efficacy assays. On the day of assay, the growth medium was carefully removed by suction and cells were washed once with 200 ⁇ of PBS, incubated in 90 ⁇ reaction media (plain DMEM media, low
  • Seq. ID. 258 73(9) 100 (1) 100 (1) 100 (0.5) a: % degradation of peptides in 24h when incubated with human plasma and values in bracket represent half-life (1 ⁇ 2), in h;
  • b % degradation of peptides in 24h when incubated with simulated gastric fluid and values in bracket represent half-life (tin), in h;
  • c % degradation of peptides in 24h when incubated with simulated intestinal fluid and values in bracket represent half-life (ti /2 ), in h;
  • the ovariectomiced (OVX) rats were used to study anti-osteoporosis activity of test compounds (cyclic short-chain peptides) in vivo.
  • the OVX rats develop osteopenia due to ovarian hormone deficiency. Osteopenia can be detected as early as 14 days post OVX, increase for the next 100 days and then stabilised (Wronski T. J., et al., Calcif. Tissue Int., 43(3), 1988, 179-183).
  • Lumbar vertebra (L5) was excised, carefully cleaned and collected in formal saline. Femur, tibia and Lumbar vertebra-5 (L5) were later processed for histomorphometric evaluation (Zhang L., et al., Tohoku J Exp Med., 1998, 186(2), 131-41; Tanizawa T., et al., Toxicol Lett., 1998,102, 399-403). Increase in serum Calcium and decrease in Phosphate levels (% change vs PTH) of selected peptides are listed in Table 5.
  • the present invention provides a method of making cyclic short chain peptides that function as an agonist of the PTH-1 receptor having different degree of affinity (1-1000 nM concentration) in UMR-106 cells.
  • the PTH-1 receptor agonistic activity was assessed by estimation of amount of cAMP released by the test compounds (in vitro).
  • OVX mice/ rat models in vivo
  • some of the cyclic short-chain peptides showed improvement in bone growth parameters thus making them ideal therapeutic candidates for the treatment and prevention of osteoporosis.
  • Novel cyclic short chain peptides of present invention showed increased stability against various proteolytic enzymes and due to increased stability and short chain length, such cyclic short chain peptides can also be delivered by oral route of administration, along with other invensive and non-invensive routes of administration.
  • the cyclic short chain peptides of the present invention can be used either alone or in combination with other anti-osteophoratic agents such as growth harmone secretagogues, Insulin-like growth factor (IGF-1), Vitamine-D analogues, estrogen, raloxifene (oestrogen receptor modulators), calcitonin, PTH-1 r agonist and the bisphosphonates (etidronate, alendronate and risedronate) for the treatment and prevention of osteoporosis.
  • IGF-1 Insulin-like growth factor
  • Vitamine-D analogues Vitamine-D analogues
  • estrogen raloxifene (oestrogen receptor modulators)
  • calcitonin calcitonin
  • PTH-1 r agonist bisphosphonates
  • novel cyclic short chain peptides of the present invention can be formulated into suitable pharmaceutically acceptable compositions by combining with suitable excipients as are well known.
  • the pharmaceutical composition containg the short is provided by employing conventional techniques.
  • the composition is in unit dosage form containing an effective amount of the active component, that is, the cyclic short-chain peptides of formula (I) either alone or combination, according to this invention.
  • the pharmaceutical composition can be prepared by known processes by combining the compound of formula (I) with suitable excipients comprising suitable excipients selected from suitable diluents, stabilizers, buffers and the like as is known in the art.
  • the quantity of active component that is, the cyclic short chain peptides of formula (I) according to this invention, in the pharmaceutical composition and unit dosage form thereof may be varied or adjusted widely depending upon the particular application method, the potency of the particular short chain peptides and the desired concentration. Generally, the quantity of active component will range between 0.5 % to 90 % by weight of the composition.
  • the cyclic short chain peptides of the present invention can be administered to mammals, preferably humans, for the treatment of a variety of conditions and disorders, including, but not limited to, treating or preventing osteoporosis, such as primary osteoporosis, endocrine osteoporosis (hyperthyroidism, hyperparathyroidism, Cushing's syndrome, acromegaly, type 1 diabetes mellitus, adrenal insufficiency), hereditary and congenital forms of osteoporosis (osteogenesis imperfecta, homocystinuria, Menkes' syndrome and Riley-Day syndrome), nutritional and gastrointestinal disorders, haematological disorders/malignancy (multiple myeloma, lymphoma and leukaemia, hemophilia-, thalassemia), osteoporosis due to immobilization, chronic obstructive pulmonary disease or rheumatologic disorders (rheumatoid arthritis, spondylitis), Osteomyelitis
  • Hypercalcemia resulting from solid tumours and hematologic malignancies idiopathic hypercalcemia and hypercalcemia associated with hyperthyroidism and renal function disorders.
  • Osteopenia following surgery induced by steroid administration and associated with disorders of the small and large intestine and with chronic hepatic and renal diseases.
  • Osteonecrosis or bone cell death associated with traumatic injury or nontraumatic necrosis associated with Gaucher's disease, sickle cell anaemia, systemic lupus erythematosus and other conditions.
  • Periodontal bone loss, Osteolytic metastasis, bone fracture healing and hyperproliferative skin disorders such as psoriasis.

Abstract

L'invention concerne de nouveaux peptides cycliques à chaîne courte présentant des propriétés pharmaceutiques bénéfiques. L'invention concerne aussi un procédé de préparation de tels peptides à chaîne courte et des compositions phamaceutiques renfermant les peptides cycliques de la présente invention. A-Z 1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z10-Z11-Z 12-Z13-Z 14-Z 15-B (I)
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Publication number Priority date Publication date Assignee Title
CN113736741A (zh) * 2021-08-18 2021-12-03 上海市食品药品检验研究院 一种用于测定特立帕肽生物学活性的克隆化细胞株
CN115925802A (zh) * 2023-02-22 2023-04-07 西南交通大学 一种具有增强抗菌活性和血清稳定性的抗菌肽及其应用

Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003009804A2 (fr) 2001-07-23 2003-02-06 The General Hospital Corporation Analogues d'hormone parathyroide (pth) a contrainte conformationnelle
WO2004067021A1 (fr) 2003-01-24 2004-08-12 Gardella Thomas J Analogues de parathormone (pth) a contrainte conformationnelle avec ponts lactame
WO2004093902A1 (fr) 2003-03-19 2004-11-04 The General Hospital Corporation Hormones parathyroidiennes contraintes de maniere conformationnelle avec des stabilisateurs en helice $g(a)
WO2005077918A1 (fr) 2004-02-11 2005-08-25 Smithkline Beecham Corporation Agonistes de pth
US20060019902A1 (en) 2004-07-19 2006-01-26 Andrea Caporale PTH-like peptides
US7153951B2 (en) 1998-10-22 2006-12-26 The General Hospital Corporation Bioactive peptides and peptide derivatives of parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP)
US20070099940A1 (en) 2004-02-11 2007-05-03 Spearing Paul K Pth agonists
US20070117157A1 (en) 2002-09-13 2007-05-24 Zajac Jeffrey D Parathyroid hormone-like polypeptides
US20070203071A1 (en) 2003-07-17 2007-08-30 Gardella Thomas J Conformationally Constrained Parthyroid Hormone (Pth) Analogs
WO2007130113A2 (fr) 2005-09-06 2007-11-15 Zelos Therapeutics, Inc. Analogues de l'hormone parathyroidienne et procedes les utilisant
WO2008068487A1 (fr) 2006-12-08 2008-06-12 Zealand Pharma A/S Peptides pth tronqués à configuration cyclique
WO2010128521A2 (fr) 2009-04-24 2010-11-11 Cadila Healthcare Limited Peptides à chaîne courte comme agoniste du récepteur de l'hormone parathyroïdienne (pth)

Patent Citations (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7153951B2 (en) 1998-10-22 2006-12-26 The General Hospital Corporation Bioactive peptides and peptide derivatives of parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP)
US20070111946A1 (en) 1998-10-22 2007-05-17 Gardella Thomas J Bioactive peptides and peptide derivatives of parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP)
WO2003009804A2 (fr) 2001-07-23 2003-02-06 The General Hospital Corporation Analogues d'hormone parathyroide (pth) a contrainte conformationnelle
US20070117157A1 (en) 2002-09-13 2007-05-24 Zajac Jeffrey D Parathyroid hormone-like polypeptides
WO2004067021A1 (fr) 2003-01-24 2004-08-12 Gardella Thomas J Analogues de parathormone (pth) a contrainte conformationnelle avec ponts lactame
WO2004093902A1 (fr) 2003-03-19 2004-11-04 The General Hospital Corporation Hormones parathyroidiennes contraintes de maniere conformationnelle avec des stabilisateurs en helice $g(a)
US20070161569A1 (en) 2003-03-19 2007-07-12 Gardella Thomas J Conformationally constrained parathyroid hormones with alpha-helix stabilizers
US20070203071A1 (en) 2003-07-17 2007-08-30 Gardella Thomas J Conformationally Constrained Parthyroid Hormone (Pth) Analogs
WO2005077918A1 (fr) 2004-02-11 2005-08-25 Smithkline Beecham Corporation Agonistes de pth
US20070099940A1 (en) 2004-02-11 2007-05-03 Spearing Paul K Pth agonists
US20060019902A1 (en) 2004-07-19 2006-01-26 Andrea Caporale PTH-like peptides
WO2007130113A2 (fr) 2005-09-06 2007-11-15 Zelos Therapeutics, Inc. Analogues de l'hormone parathyroidienne et procedes les utilisant
WO2008068487A1 (fr) 2006-12-08 2008-06-12 Zealand Pharma A/S Peptides pth tronqués à configuration cyclique
WO2010128521A2 (fr) 2009-04-24 2010-11-11 Cadila Healthcare Limited Peptides à chaîne courte comme agoniste du récepteur de l'hormone parathyroïdienne (pth)

Non-Patent Citations (64)

* Cited by examiner, † Cited by third party
Title
ANDREA C. ET AL., J.M.C., vol. 53, 2010, pages 8072 - 8079
ANDREA C. ET AL., JMC., vol. 53, 2010, pages 8072 - 79
AZURANI A. ET AL., J. BIOL. CHEM., vol. 271, 1996, pages 14931 - 14936
BETSBRUGGE J.V. ET AL., TETRAHEDRON, vol. 54, 1988, pages 1753 - 1762
BOESTEN, W.H.J. ET AL., ORG. LETT., vol. 3, no. 8, 2001, pages 1121 - 1124
CARTER P. H. ET AL., PNAS, vol. 104, no. 16, 2007, pages 6846 - 6851
CATIVIELA C.; DIAZ-DE-VILLEGAS M. D., TETRAHEDRAN ASYMMETRY, vol. 9, 1988, pages 3517 - 3599
CHOREV M., ACC. CHEM. RES., vol. 26, 1993, pages 266 - 273
COLE Z. A. ET AL., CURR. RHEUMATOL. REP., vol. 10, no. 2, 2008, pages 92 - 96
DEMPSTER D. W. ET AL., ENDOCR. REV., vol. 14, 1993, pages 690 - 709
DONNELLY D., FEBS LETTS., vol. 409, 1997, pages 431 - 436
E. ATHERTON; R.C. SHEPPARD: "Special methods in peptide synthesis, Part C", vol. 9, 1987, ACADEMIC PRESS, article "The Fluorenylmethoxycarbonyl amino protecting group'', in ''The peptides: Analysis, synthesis, Biology", pages: 1 - 38
E. GROSS; J. MEINHOFER: "The peptide synthesis, analysis, biology", vol. 1, 1979, ACADEMIC PRESS
G. BARANY; R. B. MERRIFIELD: "Special methods in peptide synthesis, Part A", vol. 2, 1980, ACADEMIC PRESS, article "The peptides: Analysis, synthesis, Biology", pages: 3 - 284
GARDELLA T. J. ET AL., ENDOCRINOLOGY, vol. 132, 1993, pages 2024 - 2030
GARDELLA T. J. ET AL., ENDOCRINOLOGY, vol. 142, 2001, pages 3068 - 3074
GARDELLA T. J. ET AL., J. BIOL. CHEM., vol. 266, 1991, pages 13141 - 13146
GARDELLA T. J. ET AL., J. BIOL. CHEM., vol. 275, 2000, pages 21836 - 21843
GARDELLA T. J. ET AL., J. BIOL. CHEM., vol. 52, 2001, pages 49003 - 49012
GOLTZMAN D., J. BONE MINER. RES., vol. 15, no. 3, 2000, pages 605 - 608
GREENSPAN S. L. ET AL., ANN. INTERN. MED., vol. 146, no. 5, 2007, pages 326 - 339
HARVEY, N. ET AL., CURR. RHEUMATOL. REP., vol. 5, no. 1, 2003, pages 75 - 81
HJORTH, S. A. ET AL., REGULATORY PEPTIDES, vol. 64, 1996, pages 70
HOARE S. R. J. ET AL., J. BIOL. CHEM., vol. 276, 2001, pages 7741 - 7753
HOARE, S. R. J., DRUG DISCOVERY TODAY, vol. 10, 2005, pages 423 - 427
J. M. STEWART; J. D. YOUNG: "Solid-phase peptide synthesis,2nd Ed.,", 1984, PIERCE CHEMICAL CO.
J. MED. CHEM., vol. 53, 2010, pages 8072 - 8079
JI T. H ET AL., J. BIOL. CHEM., vol. 273, 1988, pages 17299 - 17302
JUPPNER H. ET AL., SCIENCE, vol. 254, 1991, pages 1024 - 1026
KING D.S. ET AL., INT. J. PEPTIDE PROTEIN RES., vol. 36, 1990, pages 255 - 266
KOLAKOWSKI L. F., RECEPTOR CHANNELS, vol. 2, 1994, pages 1 - 7
KRONENBERG H. M.; BRINGHURST F. R.; NUSSBAUM S. R.; JUPPNER H.; ABOU-SAMRA A. B.: "Handbook of Experimental Pharmacology", 1993, SPRINGER-VERLAG, pages: 185 - 201
KUIJPERS G., BMJ, vol. 324, no. 7335, 2002, pages 435 - 436
LANE J. M. ET AL., CLIN. ORTHOP. RELAT. RES, vol. 372, 2000, pages 139 - 150
LANSKE B. ET AL., CRIT. REV. EUKARYOT. GENE EXPR., vol. 8, 1998, pages 297 - 320
LINDSAY R., LANCET, vol. 341, no. 8848, 1993, pages 801 - 805
LUCK M. D. ET AL., MOLECULAR ENDOCRINOLOGY, vol. 13, 1999, pages 670 - 680
M. BODANSKY; A. BODANSKY: "The practice of peptide synthesis", 1984, SPRINGER-VERLAG
MERRITT B. S., J. CELLULAR PHYSIOLOGY, vol. 152, no. 3, 2005, pages 520 - 528
NAUSSBAUM S. R. ET AL., J. BIOL. CHEM., vol. 255, 1980, pages 10183 - 10187
NEER R. M. ET AL., N. ENGL. J. MED, vol. 344, 2001, pages 1434 - 1441
NEER R. M. ET AL., N. ENGL. J. MED., vol. 344, 2001, pages 1434 - 1441
OLSON G. L. ET AL., J. MED. CHEM., vol. 36, no. 21, 1993, pages 3039 - 3049
RECKER R. R., J. CLIN. ENDOCRINOL. METAB., vol. 76, no. 1, 1993, pages 14 - 16
REID I. R., AM. J. PHYSIOL. ENDOCRINOL. METAB., vol. 253, 1987, pages E45 - E50
REIDHAAR-OLSON J. F. ET AL., MOL. CELL. ENDOCRINOLOGY., vol. 160, 2000, pages 135 - 147
RIXON R. H. ET AL., J. BONE MINER. RES., vol. 9, 1994, pages 1179 - 1189
SEEMAN E. ET AL., N. ENGL. J. MED., vol. 354, no. 21, 2006, pages 2250 - 2261
SHIMIZU M. ET AL., ENDOCRINOLOGY, vol. 142, 2001, pages 3068 - 3074
SHIMIZU M. ET AL., J. BIOL. CHEM., vol. 275, 2000, pages 21836 - 21843
SHIMIZU N. ET AL., J. BIOL. CHEM., vol. 276, 2001, pages 49003 - 49012
SLOVIK D. M. ET AL., J. BONE MINER. RES., vol. 1, 1986, pages 377 - 381
TAKASU H ET AL., BIOCHEMISTRY, vol. 38, 1999, pages 13453 - 13460
TANIZAWA T. ET AL., TOXICOL LETT., vol. 102, 1998, pages 399 - 403
TREGEAR G. W. ET AL., ENDOCRINOLOGY, vol. 93, 1973, pages 1349 - 1353
VAHLE J. L., TOXICOL. PATHOS., vol. 32, no. 4, 2004, pages 426 - 438
WHITEFILD J. F. ET AL., CALCIF. TISSUE INT., vol. 56, 1995, pages 227 - 231
WHITFIEL J. F., MEDSCAPE WOMENS HEALTH, vol. 6, no. 5, 2001, pages 7
WHITFIELD J. F. ET AL., CALCIF. TISSUE INT., vol. 58, 1996, pages 81 - 87
WHITFIELD J. F. ET AL., J. BONE MINER. RES., vol. 12, 1997, pages 1246 - 1252
WHITFIELD J. F. ET AL., TRENDS PHARMACOL. SCI., vol. 16, 1995, pages 372 - 386
WRONSKI T. J. ET AL., CALCIF. TISSUE INT., vol. 43, no. 3, 1988, pages 179 - 183
ZHANG L. ET AL., TOHOKU J EXP MED., vol. 186, no. 2, 1998, pages 131 - 41
ZUBAY, G.: "Biochemistry 2nd ed.,", 1988, MACMILLAN PUBLISHING, pages: 33

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113736741A (zh) * 2021-08-18 2021-12-03 上海市食品药品检验研究院 一种用于测定特立帕肽生物学活性的克隆化细胞株
CN115925802A (zh) * 2023-02-22 2023-04-07 西南交通大学 一种具有增强抗菌活性和血清稳定性的抗菌肽及其应用

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