MXPA97009098A

MXPA97009098A - Chemical analogues of body grease-pro with biological power increment

Info

Publication number: MXPA97009098A
Application number: MXPA/A/1997/009098A
Authority: MX
Inventors: Ibea Michel; Brazeau Paul; Abritat Thierry
Original assignee: Theratechnologies Inc
Priority date: 1995-05-26
Filing date: 1997-11-25
Publication date: 1998-10-30

Abstract

The present invention relates to FRC chimeric fatty acid analogues with increased biological potency, their application as anabolic agents and the diagnosis and treatment of growth hormone deficiencies. FRC chimeric fatty body analogues include a hydrophobic portion (glue) and can be prepared either by joining one or more hydrophobic tails to the FRC or substituting one or more amino acids for a pseudomicelar residue in the chemical synthesis of FRC. The FRC analogs of the present invention are biodegradable, non-immunogenic and exhibit improved anabolic potency at reduced dose and prolonged activity

Description

CHLORIC ANALOGUES OF GREASY-PRO BODY WITH INCREMENTED BIOLOGICAL POWER BACKGROUND OF THE INVENTION (a) FIELD OF THE INVENTION The invention relates to chimeric analogs of fatty-pro-FRC body with increased biological potency and prolonged activity, its application as anabolic agents and treatment of growth hormone deficiencies. (b) Description of the Prior Art Growth hormone (GH) or somatotropin, secreted by the pituitary gland, constitutes a family of hormones whose biological activity is fundamental for the linear growth of a young organism but also for the maintenance of integrity in your adult status HC acts directly or indirectly on peripheral organs by stimulating the synthesis of growth factors, (insulin-like growth factor-1 IGF-I) or its receptors (epidermal growth factor or FEC). The direct action of the HC is of the type denominated as anti-insulin, which favors the lipoli sis at the level of adipose tissues. Through its action on the synthesis and secretion of IGF-I (somatomedin C), HC stimulates the growth of cartilage and bones (structural growth), protein synthesis and cell proliferation in multiple peripheral organs including muscles and skin. Through its biological activity, protein synthesis and cell proliferation in multiple peripheral organs, including muscles and skin. Through its biological activity, HC participates within adults in the maintenance of a state of protein anabolism and plays a primary role in the phenomenon of tissue regeneration after trauma. The decrease of HC secretion with age, demonstrated human and animal goods, favors a metabolic change towards catabolism which initiates or participates in the aging of an organism. The loss in muscle mass, the accumulation of adipose tissue, the demineralization of the bone, the loss of capacity for regeneration of tissues after damage, which are observed in the elderly, correlates with the decrease in the secretion of HC. HC is therefore an anabolic physiological agent absolutely necessary for the linear development of children and that controls the metabolism of proteins in adults. The secretion of HC by the pituitary gland is mainly controlled by two hypothalamic peptides, somatostatin and growth hormone releasing factor (CRF). Somatostatin inhibits its secretion, whereas FRC stimulates it. Human HC has been produced by genetic engineering for approximately ten years. Until recently most of the uses of HC have been related to growth retardation in children and the uses of HC in adults have now been studied. Pharmacological uses of HC and CRF can be classified into the following three main categories. Child Development It has been shown that treatments with recombinant human growth hormone stimulate growth in children with pituitary dwarfism, renal insufficiencies, Turner syndrome and short stature. Recombinant human HC is currently marketed as a "hospice drug" in Europe and the United States for children's growth retardation caused by a deficiency of CH and for renal insufficiencies in children. The other uses are under clinical analysis research. Long-term treatment for adults and older patients A decrease in the secretion of HC causes changes in the composition of the body during aging. Preliminary studies of one year treatment with recombinant human HC reported an increase in muscle mass and skin thickness, a decrease in fat mass with a slight increase in bone density in a population of older patients. With respect to osteoporosis, recent studies suggest that recombinant human HC does not increase bone mineralization but suggests that it can prevent bone demineralization in postmenopausal women. Additional studies are currently under way to demonstrate this theory. Short-term treatment in adults and older patients In preclinical and clinical studies, it has been shown that growth hormone stimulates protein anabolism and healing in cases of burns, SI DA and cancer, in healing of wounds and bones. The HC and FRC are also intended for veterinary pharmacological uses. Both the HC and FRC stimulate growth in pigs during their fattening period favoring the deposits of muscle tissues instead of adipose tissues and increases milk production in cows and this without any unwanted side effects that could put the danger the health of the animals and without any residue in the meat or milk that is being produced. The bovine somatotropin (STB) is currently marketed in the United States. Most of the clinical studies currently conducted were conducted with recombinant HC. The FRC is considered as a second generation product intended to replace HC uses in the near future in most cases. Consequently, the use of FRC presents a number of advantages over the use of HC by itself. Physiological Advantages Growth hormone (HC) is secreted by the pituitary gland in a pulse form, since this rate of secretion is crucial for optimal biological activity. The administration of HC to correspond to this natural mode of secretion is difficult to achieve. When FRC is administered in a continuous form as a slow-release preparation or as an infusion, it increases the secretion of HC while respecting its pulsatility.

The recombinant HC that is currently marketed is the 22 kDa form whereas the FRC induces the synthesis and secretion of the pituitary gland of all the chemical isomers of HC that participates in a broader scale of biological activities. A treatment with HC results in a decreased ability of the pituitary gland to secrete endogenous growth hormone and the response of HC to FRC is decreased after said treatment. On the contrary, a treatment with CRF does not present these disadvantages, its trophic action on the pituitary gland increases the secretion capacity of this gland in normal animals and in patients with somatrotrophic insufficiency. Economic advantages The production of GF by genetic engineering is very expensive for clinical use. In particular, there are risks of contamination of this commercial preparation with material of the bacterial strain used. These bacterial contaminants can be pyrogenic or can result in patient reactions. The purification of the recombinant product is carried out following a plurality of successive chromatography steps. The criteria of drastic purity cause multiple steps of quality control. The synthesis of FRC is chemical in nature. The synthesis carried out in a solid phase and its purification is carried out in a single step using high performance liquid chromatography (HPLC). Also the amount of FRC that will be administered is much less than the amount of HC for the same resulting biological activity.

Even with all these advantages, FRC to date is not marketed as a therapeutic agent, mainly due to its chemical instability. The human FRC is a 44 amino acid peptide of the following sequence: Ala Asp Ala Me Phe Thr Asn Ser Tyr Arg Lys Val Leu Gly Gln 1 5 10 15 Leu Ser Ala Arg Lys Leu Leu Gln Asp lie Met Ser Arg Gln Gln Gly 20 25 30 Glu Ser Asn Gln Gly Arg Gly Ala Arg Ala Arg Leu-NH2 35 40 (SEQ ID NO: 1). The minimum active core is hFRC (1-29) NH2 Tyr Ala Asp Ala lie Phe Thr Asn Ser Tyr Arg Lys Val Leu Gly Gln 1 5 10 15 Leu Ser Wing Arg Lys Leu Leu Gln Asp lie Met Ser Arg 20 25 (SEQ ID NO: 2).

As for many peptides, hFRC (1-29) NH2 degrades rapidly in a serum medium and its metabolites have no residual biological activity. It has been well established that the action of enzymes, namely those of type IV dipeptidylamidopeptidase, in a blood medium, results in the hydrolysis of the binding of Ala2-Asp3 peptides from FRC. This hydrolysis results in a multitude of negative consequences that was the subject of many studies reported in the literature. Essentially, this hydrolysis leads to the formation of truncated peptides of reduced specific activity to less than 1/1000 of the biological activity.

Clinical studies with children and adults have confirmed that the natural hFRC (1 -44) N H2 or the active fragment hFRC (1 -29) N H2 are not potent enough to produce equal effects that correspond to those of recombinant HC. Many FRC analogs have been described, but all have the disadvantages of being modified FRCs that have a different amino acid sequence or that have synthetic amino acids (D series) added. These FRC analogues are potentially immunogenic and their administration to humans can cause immunotoxicity problems and potential side effects. It is well known that the binding of hydrophobic groups, such as -N Et2 at the C-terminus of a peptide sequence can result in a significantly increased specific activity. In terms of hydrophobicity, these results are contradicted by a large number of recent works such as those by Muranichi (S. Muranichi et al., 1991, Pharm. Res., 8: 649-652) which emphasize the inefficiency of the lauroyl group as a hydrophobic group used in the synthesis of small peptide analogues. Therefore, the contradictory investigations of the prior art could not direct the task of finding a more potent FRC analog using hydrophobic residues. Gaudreau and others (P. Gaudreau and others, 1992, J. Med. Chem., 35 (10): 1864-1969) describe the affinity of acetyl-, 6-aminohexanoyl-, and 8-aminooctanoyl-FRC (1-29) N H2 with the pituitary receptor of the rat . In this report, none of the tested FRC fatty acid compounds exhibited an affinity superior to that of the hFRC (1 -29) NH2 itself, and the authors concluded that "... the modifications to increase the hydrophobic character in the termination N of h FRC (1 -29) N H2 do not constitute an adequate approach to increase receptor affinity. " Coy et al., (D. H. Cow et al., 1987, J. Med. Chem., 30: 219-222.; European Patent Application published under No. 314, 866 on May 10, 1989) discloses an acetyl-FRC peptide with increased biological activity in a rat model, more particularly, in a rat anesthetized with sodium pentobarbital. The HC response in vitro was also analyzed by cultured rat pituitary cells. However, these authors do not synthesize and test fatty acid analogues-FRC with a carbon chain larger than 2 (acetyl) aggregates in the N-terminus region of the FRC. Until now, most of the FRC analogs described (including those of Gaudreau and others and those of Coy and others) have been tested in the rat model, either in vitro or in vivo. Since the FRC (1 -29) NH2 of humans and rats are notoriously different, the structure-activity relationship of FRC is different in both species. Therefore, it is not possible to extrapolate results obtained in rats for humans. The European Patent Application published under No. 320,785 on June 6, 1989, teaches FRC analogs that include a selected chain of C? -C8 alkyl; C? -C8 alkylcarbonyl; and carboxy-alkyl of d -Cβ- The European Patent Application published under No. 51 1, 003 on October 28, 1992 teaches modified amino-terminal FRC analogs having higher activity and potency with respect to FRC native or FRC analogues previously described. Consequently, it is necessary to design FRC analogues with improved anabolic potency and having a prolonged activity. This increased potency could result from a resistance to serum degradation and / or hypergonistic properties. It may be highly desirable that they be provided with FRC analogues with increased anabolic potency, while remaining biodegradable and structurally closed for natural FRC, in order to avoid immune reactions when injected chronically in humans and animals. I NVENTION DIGEST It is an object of the present invention to provide new biodegradable and non-immunogenic pro-FRC analogs with improved biological potency and prolonged activity. Another objective of the present invention is to provide pro-FRC analogs with increased anabolic potency and prolonged activity, i.e., capable of substantially raising the levels of insulin-like growth factor I (IGF-I) when administered chronically in humans And animals. Another objective of the present invention is to provide a means to return to any pro-FRC analogue that is more biologically potent and with prolonged activity.

Another objective of the present invention is to provide a method for producing active pro-FRC analogs with improved anabolic potency and prolonged activity. The present invention relates to the preparation of fatty body chimeric analogues-FRC. These chimeric analogs include a hydrophobic portion (tail) and can be prepared by joining one or more hydrophobic tails to the FRC. The pro-FRC analogs according to the present invention are characterized in that: a) These analogues have an increased biological activity; specifically, they are able to markedly increase blood levels of HC and IGF-I when administered in an animal model closely related to humans.

This feature is particularly advantageous in that it results in a reduced dose of a hyperactive compound that is administered to the patient, thereby improving the effectiveness of the treatment and reducing treatment costs. b) Both natural amino acids and hydrophobic metabolizable substances, such as fatty acids, are used for the chemical synthesis of the pro-FRC analogues. Said use of completely metabolizable natural substances is intended to avoid potential side effects, namely, in cases of multiple administrations. c) They have a high biological activity at infinitely small doses. d) They remain active for a long time, with a high biological activity. The use of fatty bodies according to the present invention results in pro-FRC analogs which overcome the drawbacks of the prior art. The pro-FRC analogs of the present invention are biodegradable, non-immunogenic and exhibit improved anabolic potency at a reduced dose and have a prolonged activity. In addition, the present invention deals with a FRC and any of its analogs, truncated or substituted. Unexpectedly, the results of the present invention showed that N-hexanoyl-, but not N-butyryl- or N-octanoyl-FRC (1-29) N H2, statistically increases IGF-I levels when administered chronically in pigs growing. These results indicate that the addition of a C4 or C8 chain in the N-terminus region of FRC produced compounds with poor biological activity when compared to N-hexanoyl-FRC (C6-FRC). Therefore, the present invention teaches that the optimum length of the carbon chain to bind to FRC in order to increase its bioactivity is from C5 to C7. This result was unexpectedly roasted in the studies published by Coy and others, which showed that the N-acetylation of FRC (addition of a C2 chain) increased its bioactivity in rats and that it did not document the activity of the compounds with a carbon chain. greater than C2. According to the method of the present invention, these analogs can be produced either by joining one or more hydrophobic tails at the N or C or FRC terminal portion or analogs thereof.

After separation and purification, the resulting modified peptide exhibits increased biological activity when administered at very low doses. According to the present invention, there is provided a chimeric fatty-body analogue FRC with enhanced biological potency of the following general formula: A1-A2-Asp-Ala-lle-Phe-Thr-A8-Ser-Tyr-Arg-Lys -Val-Leu AI5-Gln-Leu-AI8-Ala-Arg-Lys-Leu-Leu-A24-Asp-lle-A27-A28-Arq-A30 wherein: A2 is Val or Ala; A15 is Ala or Gly; A24 is Gln or His; A27 is Met, He or Nle; A28 is Ser or Asp; A30 is any amino alkyl carboxyamido-NH- (CH2) n-C0NH-, n = 1 to 12; or any amino acid sequence of 1 to 15 residues; and A1 is Tyr or His; A8 is As or Ser and A18 is Ser or Thr, where A1 is N- or O-linked by a hydrophobic tail of the general formula I: G is a carbonyl, a phosphonyl, a sulfuryl or a sulfinyl group, with a = 0 or 1; X is an oxygen atom, sulfur or an amino group (-NH-), with b = 0 or 1; The radicals R 2, R 2 and R 3 are identical or different and are selected from a hydroxyl group, a hydrogen atom and a lower linear or branched alkyl group; - (W = Y) - and - (W '= Y') represent the double bonds cis or trans - (CH = CR6) - with R5 and R6 = H or C? -C4 alkyl; with d and f = 0 or 1; R4 is a hydroxy group, a hydrogen atom or a C5-C9 alkyl; and Z is an oxygen or sulfur atom; with h = 0 to 1 with the proviso that a, b, c, d, e, f, g, and h are identical or different and that they are not all zero when R is a hydrogen, and the sum of a, b, c, d, e, f, g, and h is such that the hydrophobic tail of formula I has a linear backbone of between 5 and 8 atoms (C, O and / or S). The preferred fatty body-pro-FRC chimeric analog of the present invention is selected from the group consisting of: a) wherein A1 is Tyr or His H-alpha unique by the hydrophobic tail of formula I, wherein both a b = 1; each of d, f and h = 0; G = carbonyl; X = oxygen atom; Ri, R2, R3, R4 = hydrogen atom and the sum of c + e + g = 3, 4, 5 or 6; b) wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, wherein a = 1; each of b, d, f, and h = 0; G = carbonyl; R ,, R2, R3 and R4 = hydroxyl group and the sum c + e + g = 4, 5, 6 or 7; c) wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, wherein a = 1; each of b and h = 0; the sum of d + f = 1; G = carbonyl; Ri, R2, R3 and R4 = hydroxyl group and the sum c + e + g = 2, 3, 4, or 5; d) wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, wherein a = 1; each of b, and h = 0; the sum of d + f = 2; G = carbonyl; R ,, R2, R3 and R4 = hydroxyl group and the sum c + e + g = 0, 1, 2 or 3; e) wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, wherein a = 1; each of b, h, d and f = 0; G = carbonyl; R ,, R2, R3 and R4 = hydroxyl group and the sum c + e + g = 4, 5, 6 or 7. For the purpose of the present invention, the term "hydrophobic glue" or "Ch" is intended to mean any functionalized fatty body, such as fatty acids, fatty amines, fatty alcohols, cholesterol derivatives, etc. The term "pseudomiceal residue" or "Rp" is intended to mean any a-amino acid with side chain designed so that the residue can form or adopt a micellar structure in its switerionic form. In accordance with the present invention, there is provided a pharmaceutical formulation for inducing the release of growth hormone which comprises n active ingredient, a FRC analog of the present invention in association with a pharmaceutically acceptable carrier, excipient or diluent. In accordance with the present invention, there is provided a method for increasing the level of growth hormone in a patient comprising administering to said patient an effective amount of a FRC analog of the present invention. In accordance with the present invention, a method is provided for the diagnosis of growth hormone deficiencies in patients, which comprises administering to said patient an FRC analogue of the present invention and measuring the growth hormone response. In accordance with the present invention, there is provided a method for treating pituitary dwarfism or growth retardation in a patient comprising administering to said patient an effective amount of a FRC analog of the present invention. In accordance with the present invention, there is provided a method for treating wounds or bones in a patient, which comprises administering to said patient an effective amount of a FRC analog of the present invention. In accordance with the present invention, there is provided a method for the treatment of osteoporosis in a patient, which comprises administering to said patient an effective amount of a FRC analog of the present invention. According to the present invention, there is provided a method for improving protein anabolism (including additional protein effect) in human or animal, which comprises administering to said human or animal an effective amount of an analog of FRC of ia. present invention.

In the present invention, amino acids are identified by conventional three-letter abbreviations as indicated below, which are generally accepted in the peptide material as recommended by the IUPAC-IUB commission in biochemical nomenclature: Alanine Wing Arginine Arg Asparagine Asn Aspartic Acid Asp Cysteine Cys Glutamic Acid Glu Glycine Gly Histidine His Leucine Leu Lysine Lys Methionine Met Ornithine Orn Phenylalanine Phe Proline Pro Serine Ser Threonine Thr Tryptophan Trp Tyrosine Tyr D-Tyrosine tyr Valine Val The term "natural amino acid" means an amino acid which is present in nature or which is incorporated as a residue of free amino acids in a peptide present in nature. In addition, the abbreviation Nle is meant to mean Norleucine. Other abbreviations used are: TFA Trifluoroacetic acid; HOBt 1-Hydroxybenzothiazole DIC Di-isopropyl carbodiimide; DMF Dimethylformamide; Pip Piperidine; DMAP 4-dimethylaminopyridine; Boc t-butyloxycarbonyl Fmoc f Ioren i I met i loxicarboni lo BOP Hexachlorophosphate benzotriazo-1 -iloxitris (dimethylamino) phosphonium Me Methyl; HF Hydrofluoric acid Net3 Trieti lam ina; and TEAP Triethylammonium Phosphate (pH buffer) All peptide sequences set forth herein are described according to the generally accepted convention whereby the N-terminal amino acid is on the left and the C-terminal amino acid is on the right.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph of the effect of hFRC (1-29) NH2 analogs subcutaneously injected into serum of IGF-I pigs; Figure 2 is a curve of the effect of an intravenous injection of (4μg / kg) hFRC (1-29) NH2 and (4μg / kg) (Hexenoiltrans-3) 0 hFRC (1-29) NH2 (TT-01024) + analogue on HC of serum of pigs; Figure 3 is a graph showing the effect of various doses of hFRC (1-29) NH2 against [hexenoyl trans-3] hFRC (1-29) NH2 (TT-01024) in the area of HC below the curve over 300 minutes following administration I V. (** p <0.01 and *** p <0.001 when compared to the basal period -60 to 0 min- ); Figure 4 is a curve of the effect on a subcutaneous injection of 5 μg / kg analogs hFRC (1-29) NH2 and (5μg / kg) (Hexenoil trans-3) 0 hFRC (1-19) NH2 on HC of serum of pigs and Figure 5 is a graph showing the effect of various doses of hFRC (1-29) NH2 against [hexenoyl trans-3] ° hFRC (1-2)) NH2 (TT-01024) on the area of low HC the curve for 420 minutes after SC administration (** P < 0.01 and *** P < 0001 when compared to the basal period -60 to 0 min-). DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the use of fatty substances, namely pseudomyelic residues and / or hydrophobic tails to produce a new family of fatty-body chimeric analogues FRC while still being biodegradable and non-immunogenic. According to the present invention, the fatty-pro-FRC body analogues can be chemically synthesized: by joining one or more hydrophobic tails in the terminal portion C and / or N of FRC or one of its analogs, or incorporating one or several derivative (s) of pseudomyellar amino acids ("pseudomyellar residue") in the chemical synthesis of FRC or one of its analogues. According to the present invention, the structure of pseudomyellar residues (Rp) used as a ligation in the synthesis of FRC and analogs thereof can be represented as follows: wherein: W is a group selected from the group consisting of -CO2Q3; Q3 is a hydrogen atom, an ammonium ion, an element selected from the group consisting of the elements of group 1 a of Mendeleev's periodic table, or a functional group derived from the following fatty substances, pentenoic acids, hexenoic acids, heptenic acids or their saturated forms; Q? is a radical selected from the group consisting of alkyl, aralkyl, aryl and alkyl (Cn? H2 n + 1 + 1), where neither is a number between 1 and 8. Q, can be selected from the following list, which is provided to illustrate the invention instead of to limit its scope: P2-O- (CH2) "-; P8-NH-CO- (CH2) "-; P4-NH- (CH2) n-; CH, ° "fi" ~, Wherein, P, to P9 represents a hydrogen atom; a methyl group; a functional hydrophobic tail with an aliphatic, alicyclic or aromatic main chain which may be selected from the following list: saturated fatty acid of the general formula (CmH2mO2) m being a value between 4 and 12; or a side chain protecting group as described by Gross and Meinhofer (1981, The Peptides, vol.3, Academic Press: pages 1-341) such as P may be a benzyl group, bromo-2 benzyl, dichloro-2, 6 benzyl or tertiary butyl; P2 can be a benzyl or tertiary butyl group; P3 can be a benzyl, tertiary butyl, trityl, acetamidomethyl or benzamidomethyl group; P can be trifluoroacetyl, t-butyloxycarbonyl (Boc), benzyloxycarbonyl (Z) or fluorenylmethyloxycarbonyl (Fmoc); P5 can be a nitro group, p-methoxybenzenesulfonyl, mesitylenesulfonyl or pentamethylchroman; with the proviso that P6 is hydrogen, or that Ps and P6 can be adamantyloxycarbonyl; P7 can be a phenacyl, benzyloxymethyl or t-butoxymethyl group; P8 can be a benzohydryl, dimethoxybenzohydryl, trityl or xanthenyl group; n is a number between 0 and 6; And it is of the following general formula: Y = -A-Pz where: A is a bivalent heteroatom, preferably oxygen, sulfur, a group -NH- or a group -N (Me) -; Pz is the same as P ^ a P4 previously defined where Z is an integer between 1 to 4; and Q2 is a hydrogen atom. When Q ^ H or lower alkyl, Q2 can be any alkyl, alkoxy, alkyl, aralkyl or aryl group. Under these conditions, it has the same chemical identity as defined above for Qi. The carbon atoms on which (Q and (Q2) are attached are of configuration L or D. They are asymmetric but not when (Q?) = (Q2) or (W) = (Y). The union consists of one or more hydrophobic (Ch) non-pseudomiceal tails, the entire structure of said tails can be represented as follows: (Ch): R-KOfQ5 wherein: R is an alkyl, alkyl, aryl or aralkyl radical of branched or linear chains and can be derived from the group of metabolizable fatty substances consisting of saturated fatty acids of the general formula CmH2mO2, preferably with m being an integer between 4 and 6, mono- or poly-unsaturated fatty acids, fatty amines and alcohols; X represents a phosphorus atom, carbon or sulfur, f is an integer between 1 and 3, Q5 represents a hydrogen atom, ammonium bond, or an alkali metal ion, when F is an integer between 1 and 2, Q5 can be defined as for previous R with the condition that they have at least one of the following functions: Amino (-NH-); alcohol (-OH), thio (-SH), or acid (-XOfH); with X and f being as defined above. To better carry out the chemical binding reaction, hydrophobic tails or pseudomycelar residues functionalized under the acid form are preferably used. Under these conditions, the binding reaction is preferably carried out on a solid phase (Merriefield RB, 1963, J. Am. Chem. Soc., 85: 2149; 1954, J. Am. Chem. Soc, 85: 2149; 1964, J. Am. Chem. Soc., 86: 304) using extremely active reagents such as for example benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate known in the prior art (B. Castro et al., 1975, Tetrahedron letters, Vol. 14: 1219). The pseudomyellar residue to be bound is generally prepared by the direct action of a malonic salt, preferably a sodium salt of diethylacetamidomethyl malonate and the alkyl, alkyl, aryl or aralkyl halide in a polar solvent such as dimethylformamide. This reaction is usually followed by an alkaline acid or hydrolysis and a resolution (preferably enzymatic) of the resulting racemic mixture. Under certain conditions, the preparation of the pseudomyellar residue consists of: a) a first step; to protect in an orthogonal manner and to bind on a solid support of the resin type (M. Mergler et al., 1988, Peptides, Chemistry and biology, Proceedings of the 10th American peptide symposium, St. Louis, page 259, G. R., Marshall, Ed. , Escom, leiden), an amino acid with a functionalized side chain such as lysine, glutamic acid or aspartic acid; and b) a second step; to specifically deprotect the side chain and to bind at the free site a hydrophobic (Ch) metabolizable tail as described above. The pseudomicecular residue (Rp) is thus obtained after a separation (0.5% TFA / CH2Cl2) of the support-residue ligation, followed by the purification steps. The pseudom icelar residue can also be prepared by a selective complex formation of the acid and the function of alpha amine of a trifunctional amino acid, by the formation of complexes of mineral origin such as copper acetate. Under these conditions, the binding of the metabolizable hydrophobic tail is effected by the direct action of the complex formed and of said tail, either in its acyl halide form or in its acid or amine form in the presence of a condensing agent.

In the case where the hydrophobic tail to be bound consists of a fatty acid, the activation in view of the bond can be carried out in situ. Depending on the synthesis strategies used, the peptide binding site is released just prior to binding under traditional deprotection conditions (Gross and Meienhofer, 1981), The peptides, vol. 3, Academic Press: p. 1-341). The hydrophobic glue (Ch) or pseudomyellar residue (Rp) is condensed with the binding agent in organic solvents such as an ether (tetrahydrofuran), a halogenated aliphatic solvent (dichloromethane), a nitrile (acetonitrile) or an amide (dimethylformamide). With regard to bonding dynamics, the preferred working temperatures are between 20 and 60 ° C. The binding reaction time when hydrophobic glue is used are more and more hydrophobic, it varies inversely with temperature, but varies between 0.1 and 24 hours. As an illustrative example, the synthesis of triacyl lysine as set forth below, illustrates in a schematic manner the entire binding principle of a hydrophobic fatty acid tail. -OH + H0 * ^ - - * »« oc a- "H-Boc The general steps of the synthesis of FRC analogs were carried out by solid phase methodology on a peptide plus 9050 ™ synthesizer (Millipore Corporation, Milford, MA) using Fmoc strategy and synthesis cycles supplied by millipore. The Fmoc amino acids were supplied by Bachem California and other commercial sources. The sequential Fmoc chemistry using BOP / HOBt as a coupling methodology was applied to the starting resin of Fmoc-Pal-PEG (Millipore, catalog number: GEN 913383) for the production of C-terminal carboxyamides. The deprotections of Fmoc were achieved with a 20% solution of piperidine in DMF. After finishing the synthesis, the resin was washed well with DMF and ether before drying. The final separations of the side chain protection groups and peptide resin ligatures were carried out using a procedure supplied by Millipore consisting of the following mixture: TFA, water, phenol, tri-isopropylsilane (88: 5: 5: 2). ). The peptides were then precipitated and washed with ether before drying. Purification of reverse phase HPLC (buffer solution of pH A: TEAP 2.5, buffer solution of pH B: 80% CH3CN in A) used an? Ep & of water 4000, absorbency 214 nm, detector model 486, flow rate 50 ml / min; linear gradient generally from 25 to 60% B in 105 min) followed by a salting step (buffer solution of pH C: 0.1% TFA in H2O, buffer solution of pH D: 0.1% TFA in CH3CH / H2O 80:20) peptides in yields that amount to 10 to 30% with homogeneity greater than 97% calculated by HPLC (millennium disposal detection / photodiode &).

In accordance with the present invention, the pig was selected as a kind of test, since it is a valuable preclinical model for the development of FRC analogues. In addition, FRC (1-29) NH2 from human and porcine share a homology at 100% structure and the physiological pattern of HC secretion is almost identical in both species. further, the potency of FRC analogs was evaluated as their ability to significantly increase IGF-I blood levels rather than their acute HC release potency. In addition, it is known that the anabolic and healing effects of HC or FRC induced by HC are mediated by an increase in the synthesis and secretion of IGF-I. Therefore, the measurement of the elevation of IGF-I induced by FRC is the best indicator of the efficacy of the treatment. The present invention will be more readily understood by reference to the following examples which are given to illustrate the invention rather than to limit its scope. EXAMPLE 1 EFFECT OF REPEATED ADMINISTRATIONS OF [BUTIRIL0], [OCTANOIL0] -, [HEXANOIL °] - [HEXANOIL30], [HEXANOIL030], hFRC (1-29) NH2 AND [HEXANOIL0] hFRC (1 -44) NH2 AGAINST hFRC ( 1- 29) NH2 ON SERUM IGF-I LEVELS IN PIGS The objective of these experiments was to evaluate the potential of FRC analogs as anabolic agents. It is known that the secretion of HC or HC induced by FRC exerts its anabolic effect &; via an increase in the synthesis and secretion of insulin-like growth factor I (IGF-I), which results in elevated levels of circulating IGF-I. It has previously been shown that the intensity of the anabolic response to an analogous treatment of _FRC is proportional to the increase in IGF-I levels in pigs (Dubreuil P. et al., 10990, J. Anim. Sci., 68: 1254-1268 ). Therefore, in order to investigate the anabolic potency of the fatty acid-by-GFR analogs, their ability to increase IGF-I levels after repeated administrations of S.C. in pigs Experiment 1 26 Landrace x Yorkshire pigs castrated males (40-45 kg PC) were randomly distributed into 4 experimental groups: 1- hFRC (1-29) NH2 (20 μg / kg, n = 7) 2- [octanoyl0] hFRC (1-29) NH2 (20 μg / kg, n = 6) ) 3- [hexanoyl0] hFRC (1-29) NH2 (20 μg / kg, n = 6) 4- [butyryl] hFRC (1-29) NH2 (20 μg / kg, n = 7) Each animal was injected subcutaneously with IDB (twice a day) for 4 consecutive days. A blood sample was collected every morning before the first injection of the day and the day after the last injection, to measure the IGF-1. Experiment 2 40 Landrace x Yorkshire castrated male pigs (40-45 kg PC) were randomly distributed in 6 experimental groups: 1- Saline solution (n = 8) 2- 'hFRC (1-29) NH2 (40μg / kg, N = 8) 3- [Hexanoyl] hFRC (1-29) NH2 (10μg / kg, n = 8) 4- [hexanoyl0] hFRC (1-29) NH2 (20μg / kg, n = 8) 5 [hexanoyl0] hFRC (1-29) NH2 (40μg / kg, n = 8) Each animal was injected subcutaneously with IDB (twice a day) for 5 consecutive days. A blood sample was collected every morning before the first injection of the day and the day after the last injection to measure IGF-I. Experiment 3: 48 Landrace x Yorkshire castrated male pigs (40-45 kg PC) were randomly distributed into 6 experimental groups: 1- Saline solution (n = 8) 2- hFRC (1-44) NH 2 (30 μg / kg, N = 8) 3- [hexanoyl] hFRC (1-44) NH 2 (30μg / kg, n = 8) 4- [hexanoyl] hFRC (1-29) NH 2 (20μg / kg, n = 8) 5 [hexanoyl] hFRC ( 1-29) NH2 (20μg / kg, n = 8) 6 [hexanoyl0 '30] hFRC (1-29) NH2 (20μg / kg, n = 8) The selected doses were 30μg / kg for hFRC analogs ( 1-44) NH2 and 20 μg / kg for hFRC (1-29) NH2 analogues, which give identical doses on a molar basis. Each animal was injected subcutaneously with BID (twice a day) for 5 consecutive days. A blood sample was collected every morning before the first injection of the day and the day after the last injection to measure IGF-I. Measurements of IGF-I The levels of IGF were measured in serum of pigs by double radioimmunoassay of antibodies after the extraction of acid-acetic acid, as previously described (Abribat T. et al., 1993, J. Endocrinol., 39: 583-589). Extraction before radioimmunoassay is a necessary step to remove endogenous IGF binding proteins. Statistical Analysis In both experiments, the IFG-I data were analyzed by a repeated measurement analysis in two directions of variance, being the day and the treatment (FRC analog) sources of variation. Multiple comparison procedures were carried out (Student-Newman Keuls method). A P < 0.05 was considered as statistically significant. RESULTS Experiment 1 There was both a significant effect on the day (P = 0.0004) and a significant treatment per day interaction (P = 0.011), indicating that the increase in IGF-I levels depended on the tested analogue (Table 1). Blood samples for IGF-I measurements were collected daily before the first injection of compounds. The data are shown as mean + SEM of 6 to 7 values per group. Table 1 Effect of repeated SC injection (20μg / kg BID x 4 days) of FRC analogues on serum levels of IGF-I (IDB, 20μg / kg SC) treatment (ng / ml) (ng / ml (ng / ml) (ng / ml) (ng / ml) hFRC (1-29) NH2 252 + 26 235 + 19 263 + 16 258 + 17 262 + 24 [octanoyl °] hFRC (1-29) NH2 316 + 22 287 + 20 301 + 37 301 + 37 218 + 39 [hexanoil °] hFRC (1-29) NH2 248 + 20 281 + 28 299 + 26 319 + 22 * 342 + 213, b [butril °] hFRC (1-29) NH2 278 + 20 281 + 24 302 + 26 289 + 26 293 + 23 Treatment P = 0.42 Day P = 0.0004 Treatment per day P = 0.011 aP < 0.05 when compared to day 1 bP < 0.05 when compared to day 2 Multiple comparisons revealed that only [hexanoyl0] hFRC (1-29) NH2 produced an increase in IGF-I levels, which was significant on days 4 (29%, P <0.05) ) and 5 (38%, P <0.05).

FRC (1-29) human NH2 had no effect on IGF-I levels at the dose tested. Experiment 2 There was both a significant effect of the day (P < 0.0001) and a significant treatment per day interaction (P < 0.0001) indicating that the increase in IGF-I levels depended on the tested analogue (Table 2). Blood samples for measurements of IGF-I were collected daily before the first injection of the day. The data is shown as a measure + SEM of 8 values per group.

Table 2 Effect related to injection dose of repeated SC (BID x days) of FRC analogues on serum IGF-I levels Treatment Day 1 (day 2 Day 3 Day 4 Day Day 6 BIDSC treatment) (ng / ml) (ng / ml (ng / ml) (ng / ml) (ng / ml) (ng / ml) Saline 262 + 33 266 +30 281 + 34 293 + 30 287 + 32 289 + 33 hFRC (1-29) NG2 244 + 24 243 + 16 267 + 20 275 + 27 267 + 17 256 + 15 (40μg / kg) [hexanoil °] hFRC 303 + 22 327 + 20 337 + 25 338 + 25 366 + 37a 350 + 343 (1-29) NH2 (10μg / kg) [exanoil °] hFRC 302 + 38 341 + 37 368 + 43a 362 + 40a 362 + 45a 368 + 57a (1-29) NH2 (20μg / kg) [exanoil °] hFRC 252 + 35 275 + 32 319 + 31 to 354 + 41 to 350 + 34a 374 + 33abc (1-29) NH2 (40μg / kg) Treatment P = 0.23; Day P = 0.001 Treatment per day P = 0.0001 aP < 0.05 when compared to day 1 bP < 0.05 when compared to day 2 CP < 0.05 when compared to day 3 Multiple comparisons revealed that all three tested doses of [hexanoyl] hFRC (1-29) NH2 increased IGF-1 levels. At 10 μg / kg, IGF-I levels were they increased significantly on days 5 and 6 (from 16 to 21%, P <0.05). At 20μg / kg, they increased on days 3, 4, 5 and 6 (from 20 to 22%, P <0.05. At 40μg / kg, they increased on days 3, 4, 5 and 6 (from 27 to 48%, P <0.05) .The IGF-I serum levels remained stable in pigs treated with saline - and hFRC (1-29) NH2 Finally, a regression analysis revealed that the increase in IGF concentrations -I from day 1 to day 6 depended on the dose of [hexanoil0] hFRC (1-29) NH2 (? IGF-l = 11-9 + (2.77 * dose), r = 0.68, P <0.0001). There was both a significant effect of the day (P < 0.0001) and a significant treatment per day interaction (P < 0.0001) indicating that the increase in IGF-I levels depended on the tested analogue (Table IV). Multiple revealed that the analogues with a hexanoyl function were highly potent branched in the N-terminal region of FRC: [hexanoyl0] hFRC (1-29) NH2 significantly increased IGF-I levels on days 5 and 6 (by 28% and 31%, P <0.05) [hexanoil 0.30] hFRC (1-29) NH2 significantly increased IGF-I levels on days 4, 5 and 6 (by 32%, 35% and 43%, P < 0.05) [hexanoil0] hFRC (1-44) NH2 significantly increased IGF-I levels on days 3, 4, 5 and 6 (by 41%, 54%, 50% and 61%, P < 0.05) As previously observed for hFRC (1-29) NH2 (experiments 1 and 2), the entire length of hFRC (1-44) NH2 had little or no effect on IGF-I levels (except for a significant effect on day 5, which was not sustained on day 6). ).

Finally the binding of a hexanoyl function in the C-terminal region of hFRC (1-29) NH2 produced an analogue with increased potency when compared with hFRC (1-29) NH2 (21% increased in IGF-I levels on day 6, P <0.05), but was less potent than [hexanoyl] hFRC (1-44) NH2. Human FRC (1-29) NH2 and hFRC (1-44) NH2 were injected 20μg / kg and 30μg / kg, respectively, in order to achieve equimolar concentrations. The data shown are the mean + SEM of 8 values per group. Table 3 Effect of multiple SC injections of FRC analogues (BID x 5 days) on serum levels of IGF-I in growing pigs Treatment Day 1 (day 2 Day 3 Day 4 Day 5 Day 6 BIDSC treatment) (ng / ml) (ng / ml (ng / ml) (ng / ml) (ng / ml) (ng / ml) Sun. Saline 215 +21 215 + 28 219 + 25 226 + 28 249 + 30 234 + 24 hFRC (1-44) NG2 245 + 21 254 + 22 285 + 26 297 + 28 303 + 26 '296 + 26 (30μg / kg) [hexanoil °] hFRC 272 + 45 292 + 52 292 + 57 315 + 57 347 + 44'DC 356 + 44'bc (1-29) NH2 (20μg / kg) [hexanoil30] hFRC 297 + 30 270 + 25 287 + 24 278 + 18 276 + 20 327 + 24 (1-29) NH2 (20μg / kg) [exanoil °] hFRC 205 + 24 212 + 26 253 + 33 271 + 36 b 277 + 29'b 294 + 26'b (1-29) NH2 (20μg / kg) [hexanoil °] hFRC 241 + 30 290 + 33 340 + 41 '372 + 4?' B 362 + 46 ° 388 + 49'bc (1-44) NH2 (30μg / kg) Treatment P = 0.23, aP < 0.05 when compared to day 1 Day P = 0.001 bP <; 0.05 when compared to day 2 Treatment x day P = 0.0001 CP < 0.05 when compared to day 3 Conclusions Neither hFRC (1-29) NH2 nor hFRC (1-44) NH2 at doses ranging from 20 to 40 μg / kg were able to modulate IGF-I levels. However, the fatty acid binding gave more potent FRC and produced analogs with markedly enhanced activity in the secretion of IGF-I. The binding of fatty acids was efficient to improve the anabolic potency of both hFRC / 1-29) NH2 and hFRC (1-44) NH2. From the above results, it was concluded that the ideal fatty acid to be used is hexanoic acid or any fatty derivative of C6 and that it should preferably bind to the N-terminal region of FRC to give maximally potent analogues. EXAMPLE II Comparative effects of pro-FRC analogues in IGF-I levels in pigs This was a 5 day treatment, twice a day administration of S.C. of a single dose of each test article against saline. This experiment was carried out to compare the efficacy of aminohexanoyl) 0 hFRC (1-29) NH2, (hexyl formate) 0 hFRC (1-29) NH2, (hexenoyl trans-2) 0 hFRC (1-29) NH2 , (Hexanoyl trans-3) 0 hFRC (1-29) NH2 and (Muconoil) or hFRC (1-29) NH2 to that of (Hexanoil) 0 hFRC (1-29) NH2. All tested compounds belong to the same family of FRC analogs: they are a combination of natural FRC and natural fatty acids, designed to improve the activity of the molecule.

Identity of tested analogues: in sun, salt TT-01015 (Hexanol) or hFRC (1-29) NH2 20μg / kg TT-01021 (Aminoexanol) or hFRC (1-29) NH2 20μg / kg TT-01022 (Hexylfomiato) or hFRC (1-29) NH2 20μg / kg TT-01023 (Hexanol trans-2) 0 hFRC (1-29) NH2 20μg / kg TT-11024 (Hexanol trans-3) 0 hFRC (1-29) NH2 20μg / kg TT-01025 (Muconoil) or hFRC (1-29) NH2 20μg / kg Route and frequency of test article ADMINISTRATION: Two daily subcutaneous injections. TEST SYSTEM: Landrace x Yorkshire pigs. DESCRIPTION OF THE ANIMAL: Fifty six castrated pigs in growth weighing 35 kg at the time of purchase. RATION: Concentrated commercial feed (18% protein) offered at free demand. EXPERIMENTAL DESIGN: Fifty-six (56) pigs were randomly distributed in 7 experimental groups (n = 8 pigs per group). Each group received two administrations S.C. daily of the following treatments (volume: 3 ml, S.C. injection). group 1: sun saline 2 x / day 2 x / day group 2: TT-01015 20 μg / kg 2 x / day group 3: TT-01021 20 μg / kg 2 x / day group 4: TT-01022 20 μg / kg 2 x / day group 5: TT-01023 20 μg / kg 2 x / day group 6: TT-01024 20 H9 g 2 x / day group 7: TT-01025 20 ^ g 2 x / day The treatments were administered on day 1 to 5. Immediately before the injections, a blood sample was collected from each animal and additional blood samples were collected on day 6. The blood samples were allowed to coagulate, serum was collected by centrifugation and subjected to IG FI analysis. The results are shown in Figure 1 as D-IG F-I, which was defined as the increase in IGF-I levels from day 1 (predetermined levels) to day 6 (after 5 days of FRC administrations). Among all the tested analogues, only hexanoyl-, hexyl- hemiate-, hexenoyl trans2- and hexenoyl trans3-hFRC (1-29) N H2 significantly increased IGF-I levels over the 6-day study period, while not they did aminohexanoyl- and muconoil-hF RC (1 -29) N H2. Since it has been shown that hFRC (1 -29) NH2 is not effect at the same dose under the same conditions in the predicted analysis (see Example I), these results show that the addition of several C6 carbon chains in the N region -FRC terminal increases its bioactivity.

EXAMPLE III Power of intravenous HC release of (Hexenoil trans-3) 0 hFRC (1-29) NH2 against hFRC (1-29) NH2 in pigs This experiment was carried out to test the release potency of HC I.V. acute (Hexenoil trans 3) 0 hFRC (1-29) NH2, a pro-FRC analog, in a physiologically close human model and to compare it with that of hFRC (1-29) NH2. (Hexenoil trans-3) 0 hFRC (1-29) NH2 is a combination of natural hFRC (1-29) NH2 and natural fatty acids. This study was a single dose, multiple dose I V injection study. Identity of tested analogs: TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 0.25 μg / kg TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 1 μg / kg TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 μg / kg hFRC (1-29) NH2 0.25 μg / kg hFRC (1-29) NH2 1 μ9 k9 hFRC (1-29) NH2 4 ^ 9 g Route and frequency of test article ADMINISTRATION: Two acute intravenous injections. TEST SYSTEM: Landrace x Yorkshire pigs. DESCRIPTION OF THE ANIMAL: Fifty six castrated pigs in growth weighing 35 kg at the time of purchase. RATION: Concentrated commercial feed (18% protein) offered at free demand. EXPERIMENTAL DESIGN: Fifty-six (56) pigs (4 reserve animals) were placed a cannula (a catheter surgically implanted in a jugular vein) within one week, before the study. On days 1 and 7, the animals with cannula were randomly distributed into 7 groups (n = 4 pigs per group), group 1: Sun. Saline group 2: TT-01024 ° -25 μ9 k9 group 3: TT-01024 1 μ9 k9 group 4: TT-01024 4 ^ g group 5: hFRC (1 -29) NH2 ° 25 μg / k9 group 6: hFRC ( 1 -29) N H2 1 ^ 9 / k9 group 7: hFRC (1 -29) N H2 4 μQ kg Blood samples for pHC analysis were collected every 20 minutes from 1 hour before 5 hours after the injections of FRC, with additional samples 10 to 30 min after the injection (n = 21 samples). The blood samples were allowed to coagulate at + 4 ° C. The serum will be collected by centrifugation, stored at -20 ° C and subjected to pHC analysis. The results are illustrated in Figures 2 and 3. As shown in Fig. 2, hFRC (1-29) NH2 (4 μg / kg) induced a rapid HC release that was sustained for approximately 60 minutes after injection . In contrast, the hexenoyl trans3-hFRC (1 -29) NH2 injected at the same dose increased the levels of HC over a longer period, approximately 260 minutes. In addition, the HC response in the first 60 minutes was moderated, suggesting that this analog acts as a pro-FRC, being processed in serum in native FRC within minutes or hours after injection. As shown in Figure 3, which shows the effects of various doses of FRC and the analogue in the HC area under the curve (from 0 to 300 minutes after injection) the hFRC (1-29) NH2 produced an effect significant on the secretion of HC at 4 μg / kg, but not at 0.25 or 1 μg / kg, while hexenoiltrans3-hFRC (1-29) NH2 produced a significant response in all 3 doses tested. In conclusion, these results show that hexenoyl transns3-hFRC (1-29) NH2 is an FRC analogue with increased potency in HC secretion and suggests that it can act as a pro-FRC, being protected from enzymatic degradation in serum. EXAMPLE IV Subcutaneous HC release power of (Hexenoil trans-3) 0 hFRC (1-29) NH2 against hFRC (1-29) NH2 in pigs This experiment was carried out to test the potency of acute S.C. of (hexenoyl trans-3) hFRC (1-29) NH2, a pro-FRC analog, in a physiologically close human model and to compare it with that of hFRC (1-19) NH2. Identity of tested analogues TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 0.31 μg / kg TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 1.25 μg / kg TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 5 μg / kg TT-01024 (Hexanoyl trans-3) 0 hFRC (1-29) NH2 20 μg / kg hFRC (1-29) NH2 1.25 μg / kg hFRC (1-29) NH2 5 μg / kg hFRC (1-29) NH2 20 μg / kg Test item frequency path ADMINISTRATION: Acute subcutaneous injection. TEST SYSTEM: Landrace x Yorkshire pigs. DESCRIPTION OF THE ANIMAL: Sixty-four (64) castrated pigs in growth weighing 35 kg at the time of purchase. RATION: Concentrated commercial feed (18% protein) offered at free demand. EXPERIMENTAL DESIGN: Thirty-six (36) pigs (4 reserve animals) were placed a cannula (a catheter surgically implanted in a jugular vein) within a week, before the study. On days 1 and 7, the animals with cannula were randomly distributed into 7 groups (n = 4 pigs per group). group 1: saline group 2: TT-01024 0.31 μg / kg group 3: TT-01024 1.25 μg / kg group 4: TT-01024 5 μg / kg group 5: TT-01024 20 μg / kg group 6: hFRC (1 -29) NH2 1 25 μg / kg group 7: hFRC (1-29) NH2 5 ^ / kg group 8: hFRC (1 -29) NH2 20 μg / kg Blood samples for pHC analysis were collected every 20 minutes from 1 hour before to 7 hours after the FRC injections, (n = 25 samples). The blood samples were allowed to coagulate at + 4 ° C. The serum was collected by centrifugation, stored at -20 ° and subjected to pHC analysis. The results are shown in Figures 4 and 5. As shown in Figure 4, subcutaneous injection of 5 μg / kg of hFRC (1 -29) N H2 induced a HC response in the first 60 minutes after administration , while the same hexenoyl trans3-hFRC (1 -29) NH2 injection induced an HC response that was sustained for 240 minutes. Figure 5 illustrates the effect of several doses of the FRC tested in the HC area under the curve during the study period, that is, from 0 to 420 minutes after the injection. During this period, hFRC (1 -29) NH2 did not induce any significant HC response at any of the doses tested, while hexenoyl trans3-hFRC (1 -29) NH2 produced significant increases in HC AUC at 5 and 20 μg / kg. These results together suggest that hexenoyl trans3-hFRC (1 -29) N H2 is a highly potent HC secretor, even when administered subcutaneously. While the invention has been described in relation to specific embodiments thereof, it will be understood that it is capable of further modifications and its application is intended to cover any variation, use or adaptations of the following invention, in general, the principles of the invention. and including such deviations from the present disclosure as they fall within known or common practice within the subject matter of the invention and how it can be applied to the essential aspects set forth and as follows within the scope of the appended claims.

LIST OF SEQUENCES (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: THERATECHNOLOGIES INC. (B) STREET: 7701 - 17 eme Avenue (C) CITY: Montreal (D) STATE: Quebec (E) COUNTRY: Canada (F) POSTAL CODE (AP): H2A 2S5 (G) TELEPHONE: (514) 729-7904 (H) TELEFAX: (514) 593-8142 (ii) TITLE OF THE INVENTION: CHEMICAL ANALOGS OF FATTY BODY-PRO-FRC WITH INCREASED BIOLOGICAL POWER (iii) NUMBER OF SEQUENCES: 2 (iv) HOW IT IS READ ON THE COMPUTER. (A) TYPE OF MEDIUM: Soft disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS / MS-DOS (D) SOFTWARE: Patentln Relay # 1.0, Version # 1.30 (EPO) (vi) DATA OF PREVIOUS APPLICATION: (A) APPLICATION NUMBER: US 08 / 453,067 (B) DATE OF SUBMISSION: MAY 26, 1995 (2) INFORMATION FOR SEC ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) THREAD FORM: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETICAL: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: Tyr Ala Asp Ala Me Phe Thr Asn Being Tyr Arg Lys Val Leu Gly Gln 1 5 10 15 Leu Being Wing Arg Lys Leu Leu Gln Asp Me Met Being Arg Gln Gln Gly 20 25 30 Glu Being Asn Gln Gly Arg Gly Wing Arg Wing Arg Leu 35 40 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 29 amino acids (B) TYPE: amino acid (C) THREAD FORM: single (D) TOPOLOGY: linear (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETICAL: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Tyr Wing Asp Wing Me Phe Thr Asn Ser Tyr Arg Lys Val Leu Gly Gln 1 5 10 15 Leu Ser Wing Arg Lys Leu Leu Gln Asp Me Met Ser Arg 20 25

Claims

CLAIMS 1. A chimeric body analog of fatty-pro-FRC with increased biological potency of the following general formula: A1-A2-Asp-Ala-lle-Phe-Thr-A8-Ser-Tyr-Arg-Lys-Val-Leu - AI5-Gln-Leu-AI8-Ala-Arg-Lys-Leu-Leu-A24-Asp-lle-A27-A28-Arq-A30-R0 wherein: A1 is Tyr or His; A2 is Val or Ala; A8 is Asn or Ser; A15 is Ala or Gly; A18 is Ser or Thr; A24 is Gln or His; A27 is Met, Me or Nle; A28 is Ser or Asp; A30 is any amino acid sequence of 1 to 15 residues or absent; Ro is NH2 or NH- (CH2) n-C0NH2, with n = 1 to 12; and wherein A1 is N- or O-linked by a hydrophobic tail of the general formula I: R3 R2 R1 III R4- (Z) h- (CH) g- (W = Y1) f (CH) e- ( W = Y) d- (CH) c- (X) b- (G) a- wherein, G is a carbonyl, a phosphonyl, a sulfuryl or a sulfinyl group, with a = 0 or 1; X is an oxygen atom, sulfur or an amino group (-NH-); (W = Y) represents cis or trans (CH = CR5); (W '= Y') represents cis or trans (CH = CR6); Z is an oxygen or sulfur atom; Ri, R2 and R3, independently, are selected from a hydroxyl group, a hydrogen atom and a linear or branched C? -C6 alkyl group; R4 is a hydroxyl group, a hydrogen atom or an alkyl of Cs-Cg linear or branched; R5 and R6, independently, are a hydrogen atom or a linear or branched C? -C4 alkyl group; A is 0 or 1; B is 0 or 1; C is from 0 to 8; The sum of d + f is 1; E is from 0 to 8; G is from 0 to 8; H is from 0 to 1; Where the sum of a, b, c, d, e, f, g, and h is such that the hydrophobic tail of formula I has a linear main chain of between 5 and 7 atoms (C, O and / or S) .
2. The preferred fatty-body-pro-FRC body chimeric analog of claim 1, wherein A1 is Tyr or His H-alpha linked by the hydrophobic tail of formula I, wherein both a and b = 1; each of d, f and h = 0; G = carbonyl; X = oxygen atom; R ,, R2, R3, R4 = hydrogen atom and the sum of c + e + g = 3, 4, 5 or 6.
3. The fatty-body-pro-FRC chimeric analog of claim 1, wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, wherein a = 1; each of b, d, f, and h = 0; G = carbonyl; R ^ R2, R3 and R4 = hydroxyl group and the sum c + e + g = 4, 5, 6 or 7
4. The chimeric fatty-acid-pro-FRC analog of claim 1, wherein the sum of d + f = 1.
5. The chimeric fatty-body-by-GR analog of claim 4, wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, wherein a = 1; each of b and h = 0; the sum of d + f = 1; G = carbonyl; R1f R2, R3 and R = hydroxyl group and the sum c + e + g = 2, 3, 4, or 5.
6. The chimeric fatty-acid-pro-FRC analog of claim 5, wherein c is 0.
The chimeric fatty-body-pro-FRC analog of claim 6, wherein A30 is absent or Ser-Arg-GIn-GIn-gly-Glu-Ser-Asn-GIn-Glu-Arg-Gly-Ala-Arg -Ala-Arg-Leu 8.
The fatty-body-pro-FRC chimeric analog of claim 7, wherein A30 is absent and R0 is NH2.
The chimeric fatty-body-pro-FRC analog of claim 8, of the formula cisCH3- (CH2) 2-CH = CH-CO-Tyr, Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser -Tyr-Arg-Lys-Val-Leu-Gly-GIn-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-lle-Met-NH2 or transCH3- (CH2) 2-CH = CH- CO-Tyr, Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln- Asp-lle-Met-NH2.
10. The fatty-body-pro-FRC chimeric analog of claim 7, wherein R0 is NH2.
The chimeric fatty-body-pro-FRC analog of claim 10, of the formula cisCH3-CH2-CH = CH-CH2-CO-Tyr, Ala-Asp-Ala-lle-Phe-Thr-Asn-Ser- Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-GIn-Asp-lle-Met-Ser-Arg-GIn-GIn-Gly-Glu-Arg- Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu-NH2 or transCH3-CH2-CH = CH-CH2-CO-Tyr, Ala-Asp-Ala-lle-Phe-Thr- Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-Me-Met-Ser-Arg-Gln-Gln-Gly- Glu-Arg-Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu-NH2.
The chimeric fatty-body-pro-FRC analog of claim 1, wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of the formula I, wherein a = 1; each of b, and h = 0; the sum of d + f = 2; G = carbonyl; R ,, R2, R3 and R4 = hydroxyl group and the sum c + e + g = 0, 1, 2 or 3.
13. The fatty-metal body chimeric pro-FRC of claim 1, wherein A1 is Tyr or His N-alpha linked by the hydrophobic tail of formula I, where a = 1; each of b, h, d and f = 0; G = carbonyl; Ri, R2, R3 and R4 = hydroxyl group and the sum c + e + g = 4, 5, 6 or 7.
14. A pharmaceutical formulation for inducing the release of growth hormone comprising as an active ingredient an analogue of FRC according to claim 1, in association with a pharmaceutically acceptable carrier, excipient or diluent.
15. The use of an FRC analogue according to claim 1, for the diagnosis of growth hormone deficiencies in a patient, which comprises administering to said patient the FRC analog and measuring the response to growth hormone.
16. The use of an FRC analogue according to claim 1, for increasing the level of growth hormone in a patient.
17. The use of an FRC analogue according to claim 1 for the treatment of pituitary dwarfing or growth retardation in a patient.
18. The use of an FRC analogue according to claim 1, for the treatment of wounds or healing bones in a patient.
19. The use of an FRC analogue according to claim 1 for the treatment of osteroporosis in a patient.
20. The use of an FRC analog according to claim 1, to improve protein anabolism in human or animal. RESU MEN The present invention relates to chimeric-pro-FRC fatty acid analogs with increased biological potency, their application as anabolic agents and the diagnosis and treatment of growth hormone deficiencies. The chimeric-pro-FRC fatty-cell analogues include a hydrophobic portion (tail) and can be prepared either by joining one or more hydrophobic tails to the FRC, or by substituting one or several amino acids for a psedomicelar residue in the chemical synthesis of FRC The FRC analogs of the present invention are biodegradable, non-immunogenic and exhibit improved anabolic potency at a reduced dose and prolonged activity.