KR19990028303A - Compounds having growth hormone releasing properties - Google Patents

Abstract

Compounds of formula A-B-C-D (E) p are used to stimulate the release of growth hormone from the pituitary gland.

Description

Compounds having growth hormone releasing properties

Growth hormone is a hormone that stimulates the growth of all tissues that can grow. In addition, growth hormone has been known to cause metabolic processes, such as stimulation of protein synthesis and free fatty acid metabolism, and to cause the conversion of energy metabolism from carbohydrates to fatty acid metabolism. The deficiency of growth hormone can cause many serious medical disorders, for example, minor complications.

Growth hormone is released from the pituitary gland. The release is directly or indirectly under tight regulation of many hormones and neurotransmitters. Growth hormone release is stimulated by growth hormone releasing hormone and can be inhibited by somatostatin. In both cases, hormones are released from the hypothalamus, but their action is mediated mainly through specific receptors located in the pituitary gland. Other compounds that stimulate the release of growth hormone from the pituitary have also been described. For example, arginine, L-3,4-dihydroxyphenylalanine (L-Dopa), glucagon, vasopressin, PACAP (pituitary adenylate cyclase activating peptide), muscarinic receptor agonist and synthetic hexapeptide, GHRP Hormone releasing peptides) release endogenous growth hormones by either directly affecting the pituitary gland or by affecting GHRH and / or somatostatin release from the hypothalamus.

In disorders or conditions in which an increased level of growth hormone is required, the protein state of growth hormone requires parenteral administration. Moreover, parenteral administration is preferred because the other directly acting natural secretagogues, such as GHRH and PACAP, are high molecular weight polypeptides.

The use of short peptides to increase the level of growth hormone in mammals has already been described for example in EP 18 072, EP 83 864, WO 89/07110, WO 89/01711, WO 89/10933, WO 88/9780, WO 83 / 02272, WO 91/18016, WO 92/01711 and WO 93/04081.

The structure of the growth hormone releasing peptide or peptide derivative is important for their bioavailability as well as for their growth hormone releasing activity. It is therefore an object of the present invention to provide novel peptides with growth hormone releasing properties with improved properties compared to known peptides of this type.

Summary of the Invention

Or a pharmaceutically acceptable salt thereof.

ABCD (-E) _p

Wherein p is 0 or 1;

A is hydrogen or R ¹ - (CH ₂ ) _q - (X) _r - (CH ₂ ) _s --CO--,

q is 0 or an integer selected from the group: 1, 2, 3, 4, 5;

r is 0 or 1;

s is 0 or an integer selected from the group: 1, 2, 3, 4, 5;

R^OneIs selected from the group consisting of hydrogen, imidazolyl, guanidino, piperazino, morpholino, piperidino or N (R²) -R³, Wherein each R²And R³Is independently hydrogen or lower alkyl optionally substituted with one or more hydroxyl, pyridinyl or furanyl groups; And

when r is 1, X is _{-NH-, -CH 2 -, -CH =} CH-, -C (R 16) (R 17) -,

, Wherein each R¹⁶And R¹⁷Is independently hydrogen or lower alkyl;

B is (G) _t - (H) _u , wherein each of t and u is independently 0 or 1;

G, and H are natural L-amino acids or their corresponding D-isomers, or derivatives thereof such as 1,4-diaminobutyric acid, amino-isobutyric acid, 1,3-diaminopropionic acid, 4-aminophenylalanine, , 2,3,4-tetrahydroisoquinoline-3-carboxylic acid, 1,2,3,4-tetrahydrounharmane-3-carboxylic acid, N-methyl anthranilic acid, anthranilic acid, N- , Non-natural amino acids such as 3-aminomethylbenzoic acid, 3-amino-3-methylbutanoic acid, sarcosine, nipecotic acid or iso-nipecotic acid;

And when t and u are both 1, the amide bond between G and H is optionally replaced by Y-NR ¹⁸ -, wherein Y is -CO- or -CH ₂ - and R ¹⁸ is hydrogen, Aralkyl;

C is the formula _{-NH-CH ((CH 2)} W -R 4) and D- amino acids of -CO-, where

w is 0, 1 or 2;

R ⁴ is

Each of which is optionally substituted with halogen, lower alkyl, lower alkoxy, lower alkylamino, amino or hydroxy;

D is a D-amino acid of the formula -NR ²⁰ -CH ((CH ₂ ) _k -R ⁵ ) -CO- when p is 1, or D is -NR ²⁰ -CH ((CH ₂ ) and _l -R ⁵⁾ -CH ₂ -R ⁶ or _{^{-NR 20 -CH ((CH 2)}} m -R 5) -CO-R 6, where

k is 0, 1 or 2;

l is 0, 1 or 2;

m is 0, 1 or 2;

R < ²⁰ > is selected from the group consisting of lower alkyl or lower aralkyl;

R ⁵ is

Each of which is optionally substituted with halogen, lower alkyl, lower alkyloxy, amino or hydroxy; And

R ⁶ is piperazino, morpholino, piperidino, -OH or -N (R ⁷ ) -R ⁸ , wherein each R ⁷ and R ⁸ is independently hydrogen or lower alkyl;

E is when p is ^{1, -NH-CH (R 10)} - , and (CH _{2) v} -R ^9, wherein

v is 0 or group: 1, 2, 3, 4,

5, 6, 7, 8;

R ⁹ is selected from the group consisting of hydrogen, imidazolyl, guanidino, piperazino, morpholino, piperidino, -N (R ¹¹ ) -R ¹² ,

, Wherein n is 0, 1 or 2 and R < ¹⁹ > is hydrogen or lower alkyl,

, Wherein o is an integer selected from the group: 1,2,3,

Each R < ¹¹ > and R < ¹² > are independently hydrogen or lower alkyl, or

, Each of which is optionally substituted with an optionally halogenated, lower alkyl, lower alkyloxy, amino, alkylamino, hydroxy, or amino group and possibly from a hexapyranos or hexapyranosyl-hexapyranos

R¹⁰When p is 1, -H, -COOH, -CH₂-R¹³, -CO-R¹³or -CH₂-OH, < / RTI > wherein < RTI ID =

R ¹³ is piperazino, morpholino, piperidino, -OH or -N (R ¹⁴ ) -R ¹⁵ wherein each R ¹⁴ and R ¹⁵ is independently hydrogen or lower alkyl;

All amide bonds in formula I, except for the bond between C and D, can be independently replaced with-Y-NR ¹⁸ -, wherein Y is -CO- or -CH ₂ - and R ¹⁸ is hydrogen, Lower aralkyl;

The following compounds are excluded.

(3-Aminomethyl benzoyl) -D-2Nal-N-Me -D-Phe-Lys-NH 2,

H-Aib-His-D- 2Nal-N-Me-D-Phe-Lys-NH 2,

H-Aib-His-N- Me-D-2Nal-N-Me-D-Phe-Lys-NH 2,

3- (H-Aib-His-D-2Nal-N-Me-D-Phe-NH)

N-Me-D-Phe-NH) -2- (1-methyl-2-pyrrolidinyl)

((3R) - piperidine-carbonyl) -N-Me-D-2Nal -N-Me-D-Phe-Lys-NH 2,

(3-aminomethylbenzoyl) -D-2Nal-N-Me-D-Phe-NH)

N-Me-D-Phe-NH) -1- (1-methyl-2-pyrrolidinyl) ethane,

N-Me-D-Phe-NH) -1- (1-methyl-2-pyrrolidinyl) ethane,

N-Me-D-Phe-NH) -1- (1-methyl-2-pyrrolidinyl) ethane,

3- (H-Aib-His-N-Me-D-2Nal-N-Me-

N-Me-D-Phe-NH) -1-morpholinopropane, N-Me-

(3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D-Phe-

N-Me-D-Phe-NH) -1- (1-methyl-2-pyrrolidinyl) ethane,

2 - (((3R) -piperidine carbonyl) -D-2Nal-N-Me-D-Phe-NH) -1- (1-methyl-2-pyrrolidinyl) ethane.

The peptide derivatives of formula I can be prepared by substitution of the amide bond (-CO-NH-) by the above-designated -Y-NR ¹⁸ , such as, for example, aminomethylene (-CH ₂ -NH) And exhibits improved resistance to proteolysis by enzymes due to the presence of adjacent D-amino acids in peptide sequences optionally combined with terminal modifications. It is believed that the bioavailability of the peptide derivatives of the invention as compared to the peptides presented in the prior art is due to their small size and their resistance to combined protein degradation.

Through the above structural formulas and throughout this specification, the following terms have the following designated meanings.

The lower alkyl moiety is intended to include alkyl moieties, preferably of 1-6 carbon atoms, of a specified length in a linear, branched, or ring arrangement. Examples of linear alkyls include methyl, ethyl, propyl, butyl, pentyl, and hexyl. Examples of branched alkyls include isopropyl, sec-butyl, tert-butyl, isopentyl and isohexyl. Examples of ring alkyls include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

Said lower alkoxy moiety is intended to include an alkoxy moiety, preferably of 1-6 carbon atoms, of a specified length in a linear or branched or ring arrangement. Examples of linear alkoxy are methoxy, ethoxy, propoxy, butoxy, pentoxy and hexoxy. Examples of branched alkoxy are isopropoxy, sec-butoxy, tert-butoxy, esopentoxy and isohexoxy. Examples of ring alkoxy include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy and cyclohexyloxy.

Said lower alkylamino moiety is intended to include alkylamino moieties, preferably of 1-6 carbon atoms, of a specified length in a linear or branched or ring arrangement. Examples of linear alkylamino include methylamino, ethylamino, propylamino, butylamino, pentylamino and hexylamino. Examples of branched alkylamino are isopropylamino, sec-butylamino, tert-butylamino, isopentylamino and isohexylamino. Examples of ring alkylamino include cyclopropylamino, cyclobutylamino, cyclopentylamino and cyclohexylamino.

In the present context, the term " aryl " refers to phenyl optionally substituted with one or more C _1-6 -alkyl, C _1-6 -alkoxy, halogen, amino or aryl, naphthyl, pyridyl, 1-H- From the group consisting of pyrimidinyl, thiazolyl, imidazolyl, indolyl, pyrimidinyl, thiadiazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiophenyl, quinolinyl, pyrazinyl, or isothiazolyl Is intended to include aromatic rings such as selected carbocyclic and heterocyclic aromatic rings. Aryl is preferably phenyl, thienyl, imidazolyl, pyridyl, indolyl, quinoline or naphthyl optionally substituted with halogen, amino, hydroxy, C _1-6 -alkyl or C _1-6 -alkoxy.

Wherein the lower aralkyl moiety is composed of a lower alkyl moiety and an aryl moiety, wherein the lower alkyl moiety and the aryl moiety are as defined above.

The term " halogen " is intended to include Cl, F, Br and I.

Generic three-letter codes are used for natural amino acids such as Ala for alanine, for example.

The present invention relates to novel peptide derivatives, compositions containing them and their use for the treatment of medical disorders resulting from the deficiency of growth hormones.

In a preferred embodiment of the compounds of formula I, A is hydrogen, 3-N-Me-AMB, -3-AMB or Aib. When t is 1, in the compounds of formula I, G is preferably Ala, Gly, sarcosine, 3-aminomethylbenzoyl, R-nipecotinyl, nipecotic or isonipecotic acid, more preferably 3-aminomethyl Benzoyl, R-nipecotinyl, nipecotic or isonipecotic acid. u is 1 when, H is preferably His, Phe, Tic, Phe ( 4-NH 2), 3-Pyal, Gly, Ala, Sar, Pro, Tyr, Arg, Orn, 3- aminomethyl benzoic acid or D-Phe More preferably H is His, Phe or Ala, and most preferably H is His or Ala. In the compounds of formula I, C is preferably D-2-naphthylalanine (D-2Nal), D-1-naphthylalanine (D-1Nal), D-Phe or D-Trp, D-Phe, and most preferably N-Me-D-2Nal, D-2Nal, D-Phe or N-Me-D-Phe. In the compounds of formula I, D is preferably -NR ²⁰ -CH ((CH ₂ ) _k -R ⁵ ) -CO-, where k is preferably 1 and R ²⁰ is lower alkyl, more preferably D is D- Phe or D-2Nal. Most preferably D is N-Me-D-Phe- ol, N-Me-D-Phe , N-Me-D-2Nal- ol, N-Me-D-Phe -NH 2, N-Me-D- Phe-NH-Me or N-Me-D- (4-1) Phe-NH-Me.

P is 1 when in a compound of formula I, E is preferably _{Lys-NH 2, Ser-NH} 2, NH- (2- (1- piperazino) ethyl), NH- (3- (1- morpholino ) Propyl), NH- (2-aminoethyl), NH- (4-aminomethylbenzyl), NH- (benzyl), Lys-OH, NH- (1 -hydroxy- And most preferably E is NH- (2- (1-methyl-2-pyridin-2-yl) 3-N, N-dimethyl-aminopropyl, Lys-NH ₂ or Ser-NH ₂ ,

In the compounds of formula I, R ⁴ is preferably 2-naphthyl. R < ⁵ > is preferably phenyl. v is preferably 2-6 and R ⁹ is NH ₂ , 2-morpholinoethyl, 3-morpholinopropyl or (1-methylpyrrolidinyl) ethyl. R ¹⁰ is preferably -COOH, -CH ₂ -OH, -H, -CONH ₂ or -CON (CH ₃ ) ₂ .

Examples of the specific compounds of the present invention are as follows.

(2R) -2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me) -3- (2-naphthyl) propanol:

(3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-NH 2:

3 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D-

H-Aib-His-N- Me-D-2Nal-N-Me-D-Phe-NH 2:

(3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-Lys-NH 2:

H-Aib-Ala-D- 2Nal-N-Me-D-Phe-Lys-NH 2

H-Aib-His-D- 2Nal-N-Me-D-Phe-NH 2

2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-

(3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D-Phe-

(3-methylaminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D-Phe- NH) -N, N-dimethylaminopropane:

(3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-N-Me 2:

H-Aib-His-N-Me-D-2Nal-N-Me-D-

3-methyl-aminomethyl -Nme-D-2Nal-Nme- D-Phe-NH-CH 3

(1R) -1- (N - ((1R) -2- (4-iodophenyl) l- (methylcarbamoyl) ethyl-N-methylcarbamate -2- (2-naphthyl) ethyl) -N-methyl amide

Structure of unnatural amino acid residues:

Abbreviations used in peptide bond substitutions

The compounds of formula I can be prepared by conventional methods of solution or solid phase synthesis. For example, solid phase synthesis can be carried out substantially as described in Stewart and Young, Solid Phase Peptide Synthesis, 2nd Ed., Rockford, Illinois, USA, 1976. Solution peptide synthesis is described, for example, in substantially Bodansky et al., Peptide Synthesis, 2nd < RTI ID = 0.0 > Ed., New York, USA, 1976.

Aminoethylenes as substituents for amides are described in Y. Sasaki and DH Coy, Peptides 8 (1), 1987, pp. 119-121. Peptide derivatives containing mono-or di-hexapyranose-derivatized amino groups are substantially as described by R. Albert et al., Life Science 53 , 1993, pp. Lt; RTI ID = 0.0 > 517-525. ≪ / RTI > Examples of suitable mono- or di-hexapyranoses include glucose, galactose, maltose, lactose or cellobiose. Derivatives used as starting materials in the synthesis are commercially available and can afford suitable protecting groups when required or the starting materials used to prepare the " A " moiety in formula I can be prepared by well known methods and optionally And can be protected by the originally known method.

Abbreviations used for savers:

Pharmaceutically acceptable acid addition salts of the compounds of formula I are those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, acetic acid, phosphoric acid, lactic acid, maleic acid, phthalic acid, citric acid, glutaric acid, gluconic acid, methanesulfonic acid, salicylic acid, , Toluenesulfonic acid, trifluoroacetic acid, sulfamic acid, and fumaric acid with an inorganic or organic acid.

In another aspect, the invention relates to a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier or diluent.

Pharmaceutical compositions containing the compounds of the present invention may be prepared by conventional techniques, for example, as described in Remington's Pharmaceutical Sciences, 1985. The compositions may be formulated in conventional forms, for example, as capsules, tablets, , Suspension, patch, or topical application.

The pharmaceutical carrier or diluent used may be a conventional solid or liquid carrier. Examples of solid carriers are lower alkyl ethers of lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, stearic acid or cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipid, fatty acid, fatty acid amine, polyoxyethylene and water.

Similarly, the carrier or diluent may comprise any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or in admixture with the wax.

If the solid carrier is used for oral administration, the preparation may be purified in the form of a powder or pellet in a hard gelatin capsule, or it may be in the form of a troche or lozenge. The amount of solid carrier will vary, but will usually be from about 25 mg to about 1 g. Typical tablets which may be prepared by conventional purification techniques may contain:

core:

100 mg of the active compound (as the free compound or its salt)

Colloidal silicone dioxide (Aerosil) 1.5 mg

Cellulose, Microcrist (Avicel) 70 mg

Modified Cellulose Gum (Ac-Di-Sol) 7.5 mg

Magnesium stearate

coating:

HPMC Approximately 9 mg

^* Mywacett 9-40 T Approximately 0.9 mg

^* Film coated acylated monoglyceride used as plasticizer for.

If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injection, such as an aqueous or nonaqueous liquid suspension or solution.

For nasal or pulmonary administration, the preparation may contain a compound of formula (I) dissolved or suspended in a liquid carrier, especially an aqueous carrier, for aerosol application. The carrier may be, for example, a stabilizer such as propylene glycol, a surfactant such as a bile acid salt, polyethylene glycol, polypropylene glycol or polyoxyethylene higher alcohol ether, an adsorption enhancer such as lecithin (phosphatidylcholine) or cyclodextrin, May contain the same additive.

For transdermal administration, the preparation may be in a form suitable for patches or ionophoresis.

In general, the compounds of the present invention are formulated in unit dosage forms consisting of 0.0001-100 mg of the active ingredient together with a pharmaceutically acceptable carrier per unit dose.

The dose of the compound according to the present invention is suitably from 1 to 500 mg / day, for example, about 100 mg / dose, when administered to a patient, e.g., a human, as a medicament.

The compounds of formula I have been shown to possess the ability to release endogenous growth hormone in vivo. Thus, this compound may be used in the treatment of conditions requiring increased plasma growth hormone levels, such as in people with growth hormone deficiency or in older patients or livestock.

Accordingly, in a particular aspect, the present invention provides a pharmaceutical composition for stimulating the release of growth hormone from the pituitary gland, comprising a compound of formula I as active ingredient together with a pharmaceutically acceptable carrier or diluent, Salt. ≪ / RTI >

In an intensive embodiment, the present invention relates to a method of stimulating the release of growth hormone from the pituitary gland, comprising administering to a patient in need thereof an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof .

In a further embodiment, the invention relates to the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for stimulating the release of growth hormone from the pituitary gland.

The compounds of formula I have interesting pharmacological properties. Examples of such traits are the stimulation of the release of growth hormone from the pituitary gland, which has a similar effect or is used as a growth hormone by itself. The use of growth hormone can be summarized as follows: stimulation of growth hormone release in the elderly; Treatment of growth retardation, treatment of renal failure or dysfunction resulting from delayed growth, prevention of side effects of glucocorticoid catabolism, treatment of osteoporosis, stimulation of the immune system, promotion of wound healing, Treatment of short-lived physiological and chronic diseases associated with growth hormone deficient children, treatment of growth retardation associated with obesity and obesity, treatment of growth retardation associated with Prader-Willi syndrome and Turner syndrome; Promote the recovery of burn patients and reduce hospitalization period; Intrauterine growth retardation, skeletal formation, hypercortisolism, and Cushing's syndrome; Induction of pulsatile growth hormone release; Treatment of delayed wound healing and psychosocial deodorisation, treatment of pulmonary dysfunction and ventilator dependence, delay of protein catabolism after major surgery, delayed wound healing, Reduce cachexia and protein loss due to chronic diseases such as cancer or AIDS; Treatment of hyperinsulinemia, including Langerhans cell cytology, adjuvant therapy for induction of ovulation; Stimulation of thymus development and treatment of immunosuppressed patients to prevent age-related decline of thymus function, muscle strength, muscle mobility, maintenance of skin thickness, metabolic homeostasis, improvement in renal homeostasis in frail elderly, Stimulation of remodeling and cartilage growth, stimulation of the immune system in companion animals and treatment of aging disorders in companion animals, stimulation of growth in growth promoters and sheep in livestock.

For this indication, the dosage will vary depending on the compound of formula I used in the desired mode of administration and therapy. However, dosage levels of 0.0001 to 100 mg / kg body weight per day are generally administered to patients and animals to obtain effective release of endogenous growth hormone. Typically, dosage forms suitable for buccal or nasal administration comprise from about 0.0001 mg to about 100 mg, preferably from about 0.001 mg to about 50 mg of a compound of formula I, mixed with a pharmaceutically acceptable carrier or diluent.

The compounds of formula I may be administered in the form of a pharmaceutically acceptable acid addition salt or, where appropriate, as an alkali metal or alkaline earth metal or a lower alkyl ammonium salt. This salt form is considered to represent the same degree of free base form and roughly active.

Alternatively, the pharmaceutical compositions of the present invention may be comprised of a compound of Formula I in combination with one or more compounds exhibiting other activities, such as, for example, antibiotics or other pharmacologically active substances. It can be a secretion enhancer such as GHRP (1 or 6) or GHRH or its analog, growth hormone or its analog or hepatic hormone such as IGF-1 or IGF-2.

The route of administration will be any route that effectively transfers the active compound to the appropriate or desired site of action, such as oral, nasal, pulmonary, transdermal or parenteral, and oral routes are preferred.

Apart from the pharmaceutical use of the compounds of formula I, they will be useful as in vitro tools to investigate the regulation of growth hormone release.

The compounds of formula I will be useful as in vivo tools for assessing the ability of the pituitary gland to release growth hormone. For example, serum samples taken before and after administration of these compounds to a human can be assayed for growth hormone. The ability of the patient's pituitary gland to release growth hormone can be determined immediately by comparing growth hormone in each serum sample.

The compounds of formula I can be administered to commercially important animals in order to increase the rate and extent of growth and to increase milk production.

Pharmacological method

The compounds of formula I can be evaluated in vitro for their efficacy and efficacy in releasing growth hormone in a primary rat somatotext.

Rat primary somatotrophs can be intrinsically prepared as previously described (Chen et al., Endocrinology 1991, 129, 3337-3342 and Chen et al., Endocrinology 1989, 124, 2791-2798). In simple terms, rats are killed by scabbard. Quickly remove pituitary gland. The pituitary gland is digested with 0.2% collagenase, 0.2% hyaluronidase in Hanks balanced salt solution. The cells were resuspended in Dulbecco's modified Eagle's medium containing 0.37% NaHCO ₃ , 10% horse serum, 2.5% fetal bovine serum, 1% non-essential amino acids, 1% glutamine and 1% penicillin / streptomycin, / ml. Place 1 ml of this suspension in each well of a 24-well tray and leave for 2-3 days before performing the release experiment.

On the day of the experiment, the cells are washed twice with the medium containing 25 mM HEPES, pH 7.4. Growth hormone release was initiated by adding medium containing 25 mM HEPES and the test compound. The cultivation is carried out at 37 DEG C for 15 minutes. The growth hormone released into the medium after incubation is measured by standard RIA.

Compounds of formula I can be evaluated for their in vivo effects on growth hormone release in pentobarbital anesthetized magnetic rats as previously described. Briefly, adult male Sprague-Dawley rats are anesthetized with pentobarbital 50 mg / kg ip. The rats are injected with tracheal cannulae and catheters into the carotid and jugular veins after the rats are sufficiently anesthetized. After 15 minutes recovery, a blood sample is taken at time zero. The pituitary secretion promoter is administered iv and the arterial blood sample is placed on ice for 15 minutes and centrifuged at 12,000 xg for 2 minutes. Serum was gently withdrawn and the amount of growth hormone was determined using standard RIA.

The invention is further illustrated by the following examples which are not intended to limit the scope of the claimed invention in any way.

The compounds prepared in the following examples were separated into trifluoroacetic acid (TFA) salts.

Example 1

Preparation of 2 (R) -2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me) -3-phenylpropanol.

165.7 mg of Boc-N-Me-D-2Nal-OH and 165.2 mg of (R) -methylamino-3-phenylpropan-1-ol (Mckennon, MJ; Meyers, AI J. Org. -71) and 68.1 mg HOAt were dissolved in a mixture of 2 ml of DMF and 4 ml of DCM at 0 < 0 > C. 115 mg of EDAC was added and the mixture was stirred at 0 ° C for 1 h and then at rt for 18 h.

DCM was then removed from the mixture by addition of 50 ml of EtOAc and the resulting mixture was successively washed with 100 ml of 5% aqueous NaHCO _3, 100 ml of H ₂ O, 100 ml of 5% aqueous KHSO _{4 and} 100 ml of H ₂ O 100 ml. Dry the organic phase resulting in a Na ₂ SO ₄ and concentrated to an oil in vacuo on a rotary evaporator.

3-Boc-aminomethylbenzoic acid were dissolved in 10 ml of DCM by adding 2 drops of DMF and the mixture was converted into a symmetric anhydride by stirring with 191.6 mg of EDAC for 10 minutes.

A solution of the lyophilized 2 (R) - (HN-Me-D2Nal-N-Me) -3-phenylpropanol and 342 [mu] l DIEA in 5 ml DCM was added to the mixture and reacted for 20 h at rt. The reaction mixture was then concentrated to an oil and redissolved in 50 ml of EtOAc. This solution was successively extracted with 100 ml of 5% aqueous NaHCO _3, 100 ml of H ₂ O, 100 ml of 5% aqueous KHSO _{4 and} 100 ml of H ₂ O. The resulting organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to an oil on a rotary evaporator. The oil was then dissolved in 4 ml DCM / TFA 1: 1 and stirred. After 10 minutes the mixture was concentrated by a nitrogen stream and re-dissolved in the oil results in _{20 ml 70% CH 3 CN /} 0.03M HCl was added and a H ₂ O 480 ml.

Then the crude product is adjusted to pH 2.5 with _{4M H 2 SO 4, 0.05M (} NH 4) SO 28% CH 3 CN to 7 on a 25 mm × 250 mm column filled with 7μ C-18 silica pre-equilibrated in ₂ Lt; RTI ID = 0.0 > HPLC. ≪ / RTI >

The column in 40 ℃ for 47 minutes, 10 ml / min with _{0.05M (NH 4) SO 4,} 28% -38% CH 3 CN in gradient elution to collect the fraction containing the peptide and the 3 volumes of H ₂ O pH2.5 And applied to a Sep-Pak ^R C18 cartridge (Waters part. # 51910) equilibrated with 0.1% TFA. The peptide was eluted from the Sep-Pak ^R cartridge with 70% CH ₃ CN 0.1% TFA, diluted with water and then separated from the eluent by lyophilization.

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). Mass spectrometry was consistent with the expected structure within the experimental error of the present method (mass spectrometry ± 0.9 amu).

RP-HPLC analysis was carried out using Vydac 218TP54 4.6 mm x 250 mm 5 mu C-18 silica column (The Separations Group, Hesperia) eluted with UV detection at 214 nm and 1 ml / min at 42 ° C. Two different elution conditions were used:

A1: equilibrated with 5% CH ₃ CN in a buffer consisting of 0.1 M (NH ₄ ) ₂ SO ₄ , adjusted to pH 2.5 with 4M H ₂ SO ₄ , and eluted with 5% to 60% CH ₃ CN Lt; / RTI >

B1: column with _{5% CH 3 CN / 0.1%} TFA / H equilibrated with ₂ O and 5% for 50 min _{CH 3 CN / 0.1% TFA /} H 2 O to _{60% CH 3 CN / 0.1%} TFA / H 2 O Lt; / RTI >

Retention times using elution conditions A1 and B1 were 29.90 min and 31.52 min, respectively.

Synthesis of 3-Boc-aminomethylbenzoic acid

25 g of 3-cyanobenzoic acid was dissolved in 70 ml of 25% NH ₃ / H ₂ O and 200 ml of H ₂ O and 5 g of 10% Pd / C was added under nitrogen. Mixture was subsequently adjusted to a pH of 10.5 by addition of 12% NH ₃ / H ₂ O was hydrogenated at atmospheric pressure at rt. After absorption of approximately 41 H ₂ for 18 h, the reaction was terminated and the catalyst was removed by filtration. The filtrate was concentrated in vacuo to 20 ml and the unreacted starting material was removed by acidification with 200 ml of 1.5 M hydrochloric acid followed by extraction with ethyl acetate. The aqueous phase was concentrated to dryness and redissolved in 400 ml THF and 343 ml 1 M NaOH. A solution of 30 g of Boc-anhydride in 100 ml THF was added and the mixture was stirred overnight. The reaction mixture was then acidified to pH 3 with 1N HCl and extracted with 3 x 300 ml of EtOAc. The organic phase was evaporated to foam. The yield was 22 g.

Abbreviation:

r.t. Room temperature

EDAC: N-ethyl-N '- (3-dimethylaminopropyl) -carbodiimide hydrochloride

EtOAc: ethyl acetate

Boc: t-butyloxycarbonyl

N-Me-D-2Nal: N-methyl-D-2-naphthylalanine

DCM: dichloromethane

DIEA: Diisopropylethylamine

DMF: N, N-dimethylformamide

HOAt: 1-Hydroxy-7-azabenzotriazole

N-Me-D-Phe-o1: N-methyl-D-phenylalaninol

TFA: Trifluoroacetic acid

THF: tetrahydrofuran

Example 2

3 - ((3-aminomethylbenzoyl)) - N-Me-D-2Nal-N-Me-

Boc-N-Me-D-Phe-OH (279 mg) was dissolved in DMF (4 ml) and HOBt (168 mg) and EDAC

(230 mg) were stirred for 10 minutes. 3-Dimethylamino-1-propylamine (188 [mu] l) was added and the mixture was stirred at rt for 18 h. Then concentrated in vacuo to a 5% aqueous sodium carbonate was added to the hydrogen (50 ml) extract the resultant mixture with EtOAc (50ml) and the organic phase was dried over Na ₂ SO ₄ and an oil.

The oil was stirred with TFA / DCM 1: 1 (6 ml) at rt for 10 min. The TFA / DCM was then evaporated by a stream of nitrogen and the resulting oil dissolved in a mixture of 70% CH ₃ CN (10 ml), 1N HCl (3 ml) and water (37 ml) And lyophilized.

The product from lyophilization was dissolved in DMF (6 ml) and DCM (12 ml). To this mixture was added Boc-N-Me-D-2Nal-OH (494 mg), HOAt (204 mg) and DIEA (17 μl) under stirring and cooling to 0 ° C followed by addition of EDAC (288 mg). After stirring at rt for 18 h, DCM was evaporated by nitrogen stream and EtOAc (100 ml) was added. The mixture was concentrated in vacuo to a 5% aqueous sodium hydrogen carbonate and extracted twice with (100 ml) and water (100 ml), dried over Na ₂ SO ₄ and five days (480 mg).

The oil was stirred with TFA / DCM 1: 1 (6 ml) and rt for 10 min. The TFA / DCM was then evaporated by a stream of nitrogen and the resulting oil was dissolved in 70% CH ₃ CN (10 ml). 1N HCl (1 ml) and water was added (47 ml) the resultant mixture immediately five days (2 HCl, HN-Me- D-2Nal-N-Me-D-Phe-NH- (CH 2) 3 -N ( (CH ₃ ) ₂ ).

Half of the oil (2HCl, HN-Me-D-2Nal-N-Me-D-Phe-NH- (CH ₂ ) ₃ -N (CH ₃ ) ₂ )

(9 ml) and 2 drops of DMF and DIEA (342 [mu] l) were added.

This solution was added to a solution of Boc-3AMB-OH (503 mg) and EDA (192 mg) in DCM (5 ml) stirred at r.t. for 15 min.

After stirring for 20 hours, the reaction mixture was concentrated to an oil with nitrogen stream and stirred with 5% aqueous sodium bicarbonate (100 ml) for 15 minutes.

It is then added to EtOAc (50 ml) The organic phase was separated and concentrated in vacuo to an oil (340 mg), dried over Na ₂ SO ₄ and extracted with 5% aqueous sodium hydrogen carbonate (100ml) and water (100ml).

The oil was stirred with TFA / DCM 1: 1 (6 ml) and rt for 10 min. The TFA / DCM was then evaporated to a stream of nitrogen and the resulting oil dissolved in 70% CH ₃ CN (10 ml) and diluted with water to a final volume of 50 ml.

This crude product was purified by semi-preparative HPLC on an eight run and lyophilized using a similar procedure as described in Example 1. [

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 608.2 amu) was consistent with the expected structure (theoretical MH ⁺ : 608.8 amu) within the experimental error of this method.

The retention times of RP-HPLC using elution conditions A1 and B1 as defined in Example 1 were 25.23 min and 26.58 min, respectively.

Example 3

N-Me-D-Phe-NH) -1-N, N-dimethylaminopropane <

In Example 2, obtained as an oil 2HCl, HN-Me-D- 2Nal-N-Me-D-Phe-NH- (CH 2) 3 -

Half of N (CH ₃ ) ₂ was dissolved in DCM (9 ml) and 2 drops of DMF and DIEA (342 μl) were added.

This solution was added to a solution of Boc- (R) -nifecotic acid (459 mg) and EDAC (192 mg) in DCM (5 ml) stirred at rt for 15 min.

After stirring for 20 hours, the reaction mixture was concentrated to an oil with nitrogen stream and stirred with 5% aqueous sodium bicarbonate (100 ml) for 15 minutes.

Then extracted with EtOAc (50 ml) was added to separate an organic phase and an aqueous 5% sodium bicarbonate (100 ml) and water (100 ml) and concentrated in vacuo to dryness five days over Na ₂ SO _4.

The oil was stirred with TFA / DCM 1: 1 (6 ml) and rt for 10 min. The TFA / DCM was then evaporated to a stream of nitrogen and the resulting oil dissolved in 70% CH ₃ CN (10 ml) and diluted with water to a final volume of 50 ml.

This crude product was then purified by semi-preparative HPLC on 5 runs and lyophilized using a similar procedure as described in Example 1. The crude product

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 586.3 amu) was consistent with the expected structure (theoretical MH ⁺ : 585.8 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 25.33 min and 26.35 min, respectively.

Example 4

N-Me-D-Phe-NH) -1- (1-methyl-2-piperidinecarbonyl) -N-

Lt; / RTI >

Boc-N-Me-D-Phe-OH (279 mg) was dissolved in DMF (10 ml) and stirred with HOBt (168 mg) and EDAC (384 mg) for 10 minutes. 2- (aminoethyl) -1-methyl-pyrrolidine (290 μl) and DIEA (171 μl) were added and the mixture was stirred at rt for 20 h.

The mixture was then concentrated to an oil and dissolved in 50 ml water and lyophilized. The product was redissolved in 25 ml water and applied to a Sep-Pak ^R C18 cartridge (Waters part. # 43345) equilibrated with 0.03 N hydrochloric acid. The product was eluted from the Sep-Pak ^R cartridge with 70% CH ₃ CN in 0.03N hydrochloric acid, diluted with water and then separated from the eluent by lyophilization. The resulting material was stirred in TFA / DCM 1: 1 (6 ml) and rt for 10 min. The TFA / DCM was then evaporated to a stream of nitrogen and the resulting oil was dissolved in 70% CH ₃ CN (10 ml) and 1 N hydrochloric acid (2 ml) was added. The product was diluted with water (50 ml) and then isolated by lyophilization.

The resulting material was dissolved in DMF (3 ml) and added with Boc-N-Me-D-2Nal-OH (329 mg), HOAt (136 mg), EDAC (230 mg) and DIEA (171 μl) Lt; / RTI > for 18 hours. EtOAc (50 ml) was then added and the mixture was extracted with 5% aqueous sodium bicarbonate (50 ml), 5% aqueous sodium hydrogen sulfate (50 ml), triturated (50 ml). The organic phase was dried over sodium sulfate and concentrated in vacuo with oil.

The oil was stirred with TFA / DCM 1: 1 (6 ml) and rt for 10 min. Then TFA / DCM to evaporate in a stream of nitrogen and dissolve the resulting oil in a _{70% CH 3 CN (10 ml} ) was added and 1N hydrochloric acid (3 ml). The product was diluted with water (50 ml) and then lyophilized.

286 mg of this lyophilized product was dissolved in DCM (15 ml) and DIEA (171 [mu] l). This solution was added to a solution of Boc- (R) -nifecotric acid (459 mg) and EDAC (192 mg) in DCM (10 ml) stirred at rt for 25 min.

After stirring for 20 h, the reaction mixture was concentrated to an oil with a stream of nitrogen and then redissolved in EtOAc (100 ml) and washed with 5% aqueous sodium bicarbonate (50 ml), 5% aqueous sodium hydrogen sulfate (50 ml) 50 ml). The organic phase was dried over sodium sulfate and concentrated in vacuo to an oil.

The oil was stirred with TFA / DCM 1: 1 (6 ml) and rt for 10 min. The TFA / DCM was then evaporated to a stream of nitrogen and the resulting oil dissolved in 70% CH ₃ CN (10 ml) and diluted with water to a final volume of 50 ml.

This crude product was then purified by semi-preparative HPLC on a 3 run and lyophilized using a similar procedure as described in Example 1. [

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 612.2 amu) was consistent with the expected structure (theoretical MH ⁺ : 612.39 amu) within the experimental error of this method.

The RP-HPLC retention time using elution condition A1 as defined in Example 1 was found to be 25.80 minutes.

Example 5

(2R) -2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me) -3- (2-naphthyl) propanol

(R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid methyl ester

(5.0 g; 16.4 mmol) was dissolved in dry DMF (50 ml). Iodomethane (6.2 ml; 98.4 mmol) and silver (I) oxide (13.3 g; 57.4 mmol) were added and the mixture was stirred overnight. The reaction mixture was filtered and the filtrate was extracted with methylene chloride (200 ml). The organic phase was washed with potassium cyanide (2 x 50 ml; 5%) and water (3 x 75 ml). The organic phase was dried (MgSO ₄₎ and the solvent was removed in vacuo. The residue was chromatographed (silica, 5 x 40 cm) using ethyl acetate and heptane (1: 2) as the eluent to give (R) -2- (N-tert-butoxycarbonyl- To obtain 4.98 g of 3- (2-naphthyl) propionic acid methyl ester.

_{^{1 H-NMR (CDCl 3)}} : 1.30, 1.35 (2 s, 9H); 2.71, 2.75 (2s, 3H); 3.19, 3.47 (2, m, 2H); 3.74, 3.77 (2s, 3H); 4.65, 5.05 (2 dd, 1 H); 7.29-7.82 (m, 7H) (mixture of rotamers)

(R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid

(R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (2-naphthyl) propionic acid

Methyl ester (21.73 g; 65.57 mmol) was dissolved in 1,4-dioxane (200 ml) and water (20 ml) was added. The reaction mixture was cooled in an ice bath and lithium hydroxide (1.73 g; 72.13 mmol) was added. After 15 minutes, water (140 ml) was added and the reaction mixture was then stirred at room temperature for a further 3 hours. Ethyl acetate (400 ml) and water (300 ml) were added and the pH was adjusted to 2.5 with 1 M sodium hydrogen sulfate (110 ml). The phases were separated and the aqueous phase was extracted with ethyl acetate (200 ml). The combined organic phase was washed with water (300 ml), dried (MgSO ₄₎ and by removing the solvent in vacuo (R) -2- (N-tert- butoxycarbonyl -N- methylamino) -3- (2 -Naphthyl) propionic acid (20.1 g).

≪ ¹ > H-NMR (DMSO): 1.18, 1.21 (2s, 9H); 2.62, 2.66 (2s, 3H); 3.11, 3.58 (m, 2H); 4.75, 4.90 (2 dd, 1 H); 7.48-7.88 (m, 7H); 1.85 (s, (br), 1H) (mixture of rotamers).

(R) -2-formylamino-3- (2-naphthyl) propionic acid

(18.11 g; 84.14 mmol) was dissolved in formic acid (204 ml) and acetic anhydride (70 ml) was added dropwise. The reaction mixture was heated to 55 < 0 > C and stirred at room temperature for 3 1/2 hours. Cooling water (70 ml) was added dropwise and the mixture was stirred at 0 ° C for 20 minutes. The reaction mixture was filtered and washed with cold water (20 ml) to obtain 20.26 g of (R) -2-formylamino-3- (2-naphthyl) propionic acid.

^{1 H-NMR (DMSO):} 3.05 (dd, 1H); 3.27 (dd, 1 H); 4.64 (m, 1 H); 7.48-7.87 (m, 7H); 7.95 (s, 1 H); 8.45 (d, 1 H); 12.9 (s, br).

(R) -methylamino-3- (2-naphthyl) propan-1-ol

(4.37 g; 18 mmol) was dissolved in dry tetrahydrofuran (100 ml) and sodium borohydride (1.6 g; 43.2 mmol) was added to the solution of (R) -2-formylamino- . Iodine (4.57 g; 18 mmol) was dissolved in dry tetrahydrofuran (40 ml) and added dropwise to the reaction mixture below 40 < 0 > C. After the addition, the reaction mixture was heated to reflux for 12 hours. Potassium hydroxide (50 ml; 20%) was added. The aqueous phase was extracted with methyl tert-butyl ether (4 x 50 ml). The combined organic layers were washed with a saturated aqueous sodium chloride (150 ml), dried (MgSO ₄₎ and the solvent was removed in vacuo. The residue was chromatographed (silica; 5 x 40 cm) using DCM / methanol / ammonia (100: 10: 1) to give (R) -methylamino- 3- (2-naphthyl) propan- g.

_{^{1 H-NMR (CDCl 3)}} : 3.43 (s, 3H); 2.88-3.05 (m, 3 H); 3.10 (s (br), 2H), 3.42 (dd, 1H); 3.69 (dd, 1 H); 7.30-7.82 (m, 7H).

N - {(1 R) -1 - [N - ((1 R) -2-hydroxy-1 - ((2-naphthyl) methyl) ethyl) -N- methylcarbamoyl] -2- Yl) ethyl} -N-methylcarbamic acid tert-butyl ester.

(2-naphthyl) propionic acid (0.55 g; 1.67 mmol) and (R) -methylamino-3- (2-naphthyl) (0.38 g; 2.00 mmol) was dissolved in methylene chloride (15 ml) and dimethylformamide (7.5 ml). The reaction mixture was cooled in an ice bath. (0.24 g; 2.09 mmol) and N- (3-dimethylaminopropyl) -N-ethylcarbodiimide hydrochloride (0.38 g; 2.0 mmol) were added to a solution of 1-hydroxy-7-azabenzotriazole The reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo. Ethyl acetate (200 ml) was added and the organic solution was washed with water (100 ml), sodium hydrogen carbonate / sodium carbonate (pH 9) (75 ml), sodium hydrogen sulfate (75 ml; 10% were (MgSO _4). The solvent was removed in vacuo and the residue was chromatographed (silica, 2 x 45 cm) using ethyl acetate to give N - {(1R) -1- (N - [(1R) -2- (2-naphthyl) methyl) ethyl) -N-methylcarbamoyl] -2- (2-naphthyl) ethyl} -N-methylcarbamic acid tert-butyl ester 0.25 g.

^& Lt; ¹ > H-NMR (DMSO): 0.80-1.99 (some s, 9H); 2.45-4.20 (m, 12H); 4.70-5.12 (m, 2H) (mixture of the selected peaks, rotamers)

(2R) -N - ((1 R) -2-hydroxy-1 - ((2-naphthyl) methyl) ethyl) -N-methyl-2-methylamino-3- (2-naphthyl) propionamide.

N - {(1R) -1- (N - [(1R) -2-hydroxy-1 - ((2-naphthyl) methyl) ethyl) -N- methylcarbamoyl] -2- Yl) ethyl} -N-methylcarbamic acid tert-butyl ester (0.25 g; 0.475 mmol) was dissolved in DCM (3 ml). Trifluoroacetic acid (1 ml) was added and the reaction mixture was stirred for 20 minutes. The solvent was removed in vacuo. DCM (5 ml) was added, removed in vacuo and repeated. The residue was dissolved in methanol (5 ml). Sodium bicarbonate / sodium carbonate (5 ml; pH 9) was added and the solution was extracted with ethyl acetate (2 x 10 ml). The organic phase was dried (MgSO ₄ ) and the solvent was removed to give (2R) -N - ((1 R) -2-hydroxy-1 - ((2-naphthyl) methyl) ethyl) -3- (2-naphthyl) propionamide (0.22 g).

_{^{1 H-NMR (CDCl 3)}} : 1.70, 2.37, 2.45, 2.93 (4 s, 6H); 2.56-3.05 (m, 2H); 3.52-3.85 (m, 7H); 4.25, 4.97 (2 m, 1 H); 6.86-7.78 (m, 14H) (selected peak, mixture of rotamers)

(2R) -2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me) -3- (2-naphthyl) propanol

3-Boc-aminomethylbenzoic acid (502.6 mg) was dissolved in DCM (6 ml) and then converted to symmetric anhydride by stirring with EDAC (191.6 mg) for 15 min.

Methyl-2-methylamino-3- (2- (2-fluorophenyl) methyl) ethyl) -N- Naphthyl) propionamide (200 mg) was added to the mixture and reacted at rt for 20 hours.

The reaction mixture was then concentrated to an oil and redissolved in EtOAc (100 ml). This solution was successively extracted with 5% aqueous NaHCO ₃ (2 × 50 ml), 5% aqueous KHSO ₄ (2 × 50 ml) and H ₂ O (2 × 50 ml). Dry the organic phase resulting in a Na ₂ SO ₄ and concentrated to an oil in vacuo on a rotary evaporator. The oil was then dissolved in DCM / TFA 1: 1 (6 ml) and stirred. The mixture was concentrated after 10 minutes in a nitrogen stream and re-dissolved in the oil results in _{70% CH 3 CN / 0.1%} TFA (5 ml) and diluted with water to a volume of 100ml.

This crude product was then purified by semi-preparative HPLC on both runs and lyophilized using a similar procedure as described in Example 1.

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 559.5 amu) was consistent with the expected structure (theoretical MH ⁺ : 560.72 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were found to be 33.07 min and 34.63 min, respectively.

Example 6

H-Aib-His-D- 2Nal-N-Me-D-Phe-NH 2

Peptides were synthesized according to the Fmoc method on an Applied Biosystems 413A peptide synthesizer on a 0.22 mmol scale using the manufacturer-provided FastMoc UV protocol using HBTU-mediated coupling in NMP and UV monitoring of deprotection of the Fmoc protecting group. The starting resin used for the synthesis was #: D-1675 (427) mg from Bachem Feinchemikalien AG, Bubendorf, Switzerland, which was reacted with Fmoc-2,4-dimethoxy- Carboxymethyloxy) -benzhydryl-amine. ≪ / RTI > The displacement capacity was 0.55 mmol / g. The protected amino acid derivatives used were Fmoc-N-Me-D-Phe-OH, Fmoc-D-2Nal-OH, Fmoc-His (Trt) and Fmoc-Aib-OH. Coupling of Fmoc-N-Me-D-Phe-OH was performed with double coupling. At room temperature and by a mixture of the peptide after synthesis 8 ml TFA, 600 mg phenol, 200μl ethanedithiol, thioanisole and 400μl 400μl H ₂ O was stirred 180 minutes were cut to 750 mg of the peptide from the resin. The crop mixture was filtered and the filtrate was concentrated to about 2 ml with nitrogen stream. The crude peptide was precipitated from this oil with 50 ml diethyl ether and washed twice with 50 ml diethyl ether.

The crude peptide was dried and purified by semi-preparative HPLC in one run and lyophilized using a similar procedure as described in Example 1.

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 598.5 amu) was consistent with the expected structure (theoretical MH ⁺ : 598.73 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 24.68 min and 25.58 min, respectively.

Example 7

H-Aib-His-D- 2Nal-N-Me-D-Phe-Ser-NH 2

This compound was synthesized using a similar procedure as described in example 6. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 685.6 amu) was consistent with the expected structure (theoretical MH ⁺ : 685.81 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 24.42 min and 25.92 min, respectively.

Example 8

(3-Aminomethyl benzoyl) -D-2Nal-N-Me -D-Phe-NH 2

This compound was synthesized using a similar procedure as described in example 6. Only the coupling of Fmoc-D-2Nal-OH was performed using HATU as activator. The H-N-Me-D-Phe resin (0.23 mmol) was coupled with 1 mmol Fmoc-D-2Nal-OH in the presence of DIEA (2 mmol) using 1 mmol HATU for 150 min.

The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 511.2 amu) was consistent with the expected structure (theoretical MH ⁺ : 509.6 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 30.73 min and 32.47 min, respectively.

Example 9

(4-piperidine-carbonyl) -D-2Nal-N-Me -D-Phe-NH 2

This compound was synthesized using a similar procedure as described in example 8. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 486.8 amu) was consistent with the expected structure (theoretical MH ⁺ : 487.6 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 27.03 min and 28.48 min, respectively.

Example 10

((3R) -3- piperidin-carbonyl) -D-2Nal-N-Me -D-Phe-NH 2

This compound was synthesized using a similar procedure as described in example 8. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 486.9 amu) was consistent with the expected structure (theoretical MH ⁺ : 487.6 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 28.03 min and 29.50 min, respectively.

Example 11

(3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-NH 2

This compound was synthesized using a similar procedure as described in Example 8 except that the final residue was introduced using symmetrical anhydride coupling. Boc-3-aminomethylbenzoic acid (251 mg) was stirred with EDAC (96 mg) in DCM for 15 min. The resin (429 mg) was then added and stirring was continued for 18 h. The other exception was that the time used to cut the peptide from the resin was reduced to 60 minutes. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 522.9 amu) was consistent with the expected structure (theoretical MH ⁺ : 523.6 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were found to be 28.83 min and 30.13 min, respectively.

Example 12

H-Aib-His-N- Me-D-2Nal-N-Me-D-Phe-NH 2

This compound was synthesized using a similar procedure as described in example 8. Here, Fmoc-N-Me-D-2Nal-OH and Fmoc-His (Trt) were coupled using HATU and the time used to cut the peptide from the resin was reduced to 60 minutes. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 612.3 amu) was consistent with the expected structure (theoretical MH ⁺ : 612.8 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were found to be 24.33 min and 26.20 min, respectively.

Example 13

(3-Aminomethyl benzoyl) -D-Phe-N-Me -D-Phe-Lys-NH 2

This compound was synthesized using a similar procedure as described in example 8. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 587.2 amu) was consistent with the expected structure (theoretical MH ⁺ : 586.74 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 21.13 min and 22.60 min, respectively.

Example 14

(3-aminomethyl-benzoyl) -N-ME-D-Phe -N-Me-D-Phe-Lys-NH 2

This compound was synthesized using a similar procedure as described in example 8. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 601.6 amu) was consistent with the expected structure (theoretical MH ⁺ : 601.77 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were found to be 20.40 min and 21.70 min, respectively.

Example 15

((3R) -3- piperidin-carbonyl) -N-ME-D-Phe -N-Me-D-Phe-Lys-NH 2

This compound was synthesized using a similar procedure as described in example 11. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 579.4 amu) was consistent with the expected structure (theoretical MH ⁺ : 579.8 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were found to be 19.88 min and 21.20 min, respectively.

Example 16

H-Aib-His-N- Me-D-Phe-N-Me-D-Phe-Lys-NH 2

This compound was synthesized using a similar procedure as described in example 12. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 690.6 amu) was consistent with the expected structure (theoretical MH ⁺ : 690.9 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 15.71 min and 17.82 min, respectively.

Example 17

((3R) -3- piperidin-carbonyl) -N-Me-D-2Nal -N-Me-D-Phe-NH 2

This compound was subjected to a similar procedure as described for Example 11 using Fmoc-N-Me-D-Phe-OH, Fmoc-N-Me-D-2Nal-OH and Boc- , Where Fmoc-N-Me-D-2Nal-OH and Boc- (R) -nifecotic acid were coupled using HATU. The resulting final product was characterized by analytical RP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ⁺ : 500.7 amu) was consistent with the expected structure (theoretical MH ⁺ : 501.7 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 28.18 min and 29.55 min, respectively.

Example 18

H-Aib-Ala-D- 2Nal-N-Me-D-Phe-Lys-NH 2

This compound was synthesized using a similar procedure as described in example 6. The resulting final product was characterized by analytical HP-HPLC (retention time) and plasma desorption mass spectrometry (molecular mass). The observed molecular mass (MH ^+: 660.7 amu) was consistent with the expected structure (theoretical MH ^+: 660.8 amu) within the experimental error of this method.

The RP-HPLC retention times using elution conditions A1 and B1 as defined in Example 1 were 25.63 min and 26.75 min, respectively.

Example 19

H-amino-3- methyl-benzoyl -N-Me-D-2Nal- N-Me-D-Phe-NH-CH 3

(62 mg, 0.458 mmol) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (97 mg, 0.458 mmol) were added to a solution of Boc-3AMB- mg, 0.504 mmol) were dissolved in DCM (8 ml) and DMF (1 ml) and stirred for 15 min. To a solution of N-methyl-2-methylamino-N - ((1 R) -1- (methylcarbamoyl) -2-phenylethyl) -3- (2-naphthyl) propionamide 185 mg, 0.458 mmol), diisopropylethylamine (80 ml, 0.458 mmol) was added, and the mixture was stirred for 20 hours.

The organic phase was washed with sodium bicarbonate (50 ml, 5%), H ₂ O (50 ml) and saturated NaCl / H ₂ O (50 ml), dried over sodium sulfate and evaporated in vacuo. The residue was dissolved in DCM (2 ml) and treated with TFA (2 ml) for 10 min. The volatiles were removed by N ₂ airflow. The residue was dissolved in 20% MeCN 50 ml and diluted to 500 ml with H ₂ O.

Cleavage HPLC

10 ml / min, run 5, 30-40% MeCN / 0.1M (NH 4) 2 SO 4, pH2.5

Detection 276 nm, Sep-Pals, 70% MeCN / 0.1% TFA, freeze-dried

PD-MS Theoretical value 536.7

Found 535.7 ± 1

HPLC A1 r _t 31.20 min

B1 r _t 36.35 min

Example 20

H-Aib-His-N-Me-D-2Nal-N-Me-D-Phe-NHMe

(BACHEM B-1570) and 1-hydroxylazobenzotriazole (338 mg, 2.48 mmol) were dissolved in 9 ml of DMF and treated with 0-4 < RTI ID = 0.0 > C and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (475 mg, 2.48

mmol) were added. The reaction mixture was stirred at < RTI ID = 0.0 > 0-4 C < / RTI > To a solution of N-methyl-2-methylamino-N - ((1 R) -1- (methylcarbamoyl) -2-phenylethyl) -3- (2-naphthyl) propionamide dissolved in methylene chloride (18 ml) (500 mg, 1.24 mmol) was cooled to 0-4 째 C, stirred at 0-4 째 C for 1 hour, and diisopropylethylamine (0.425 ml, 2.48 mmol) was added. The temperature of the mixture was slowly raised to room temperature and the mixture was stirred for 72 hours. DCM was evaporated to a stream of N ₂ and to this mixture was added 100 ml ethyl acetate and washed with sodium bicarbonate (2 x 100 ml, 5%) and sodium hydrogen sulphate (100 ml, 5%). The phases were separated and the organic phase was dried over sodium sulfate and evaporated in vacuo. The residue was dissolved in DMF (8 ml), treated with piperidine for 15 min, diluted with H ₂ O (100 ml) and quenched with acetic acid (1.5 ml). Acetonitrile was added and the mixture was diluted with H ₂ O to 250 ml. A clean solution was applied to 10 g "Seppaks"# Water, washed with H ₂ O / 0.03M HCl and eluted with 50 ml of 35% MeCN / 0.03M HCl. Diluted to 200 ml with H ₂ O and lyophilized.

(506 mg, 3.72 mmol) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (713 mg, , 3.72 mmol) to DMF (6 ml) was dissolved was added to the HL-His _{(trityl) -NMeD2Nal-NMeDPhe-NHCH 3,} 2 HCl dissolved in DCM (12 ml) after 15 minutes, diisopropylethylamine and the ( 0.637 ml) was added and the mixture was stirred for 72 hours. The DCM was evaporated to a stream of N ₂ and to the mixture was added 100 ml ethyl acetate and washed with sodium bicarbonate (2 x 50 ml, 5%) and potassium hydrogen sulphate (50 ml, 5%). The phases were separated and the organic phase was dried over sodium sulfate and evaporated in vacuo. The residue was dissolved in DCM (6 ml), cooled to 0-4 [deg.] C and treated with TFA (6 ml) for 10 min at 0-4 [deg.] C. The volatiles were removed by N ₂ airflow. The oil residue was dissolved in 35 ml of 70% acetonitrile, diluted to 50 ml with H ₂ O, 10 ml of concentrated hydrochloric acid (12 mol) was added, and the mixture was stirred for 72 hours. The mixture was diluted with H ₂ O to 200 ml, neutralized with solid sodium carbonate and finally diluted to 400 ml with H ₂ O.

Cleavage HPLC

PD-MS, calculated: 550.7, found 550.1

HPLC A1 r _t 31.75 min

B1 r _t 36.15 min

Example 21

3-methyl-amino-methyl-benzoyl -N-Me-D-2Nal- N-Me-D-Phe-NH-CH 3

(338 mg, 2.48 mmol) and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (478 mg, 2.48 mmol) were added to a solution of Boc-NMe3AMB- mg, 2.48 mmol) were dissolved in DMF (6 ml) and stirred for 15 minutes. To a solution of N-methyl-2-methylamino-N - ((1 R) -1- (methylcarbamoyl) -2- phenylethyl) -3- (2-naphthyl) propionamide dissolved in methylene chloride (12 ml) (500 mg, 1.24 mmol), followed by diisopropylethylamine (0.425 ml, 2.48 mmol). The mixture was stirred for 20 hours.

The DCM was evaporated to a stream of N ₂ and to the mixture was added 75 ml of ethyl acetate and washed with sodium bicarbonate (2 x 50 ml, 5%) and potassium hydrogen sulphate (50 ml, 5%). The phases were separated and the organic phase was dried over sodium sulfate and evaporated in vacuo. The residue was dissolved in 10 ml of methylene chloride, cooled to 0-4 [deg.] C and treated with TFA (10 ml) at 0-4 [deg.] C for 10 min. The volatiles were removed by N ₂ airflow. The oil residue was 70% MeCN / 0.1% TFA were dissolved in 25 ml, diluted with 600 ml with H ₂ O.

Cleavage HPLC

Large column, 40 ml / min, 8 run _{28-40 & P11 (NH 4) 2} SO 4, 276 nM.

Seppak, freeze-dried

PD-MS, calculated: 550.7, found 550.1

HPLC A1 r _t 31.75 min

B1 r _t 36.15 min

Example 22

(1R) -1- (N - ((1 R) -2- (4-iodophenyl) -1- (methylcarbamoyl) ethyl) -N- methyl Carbamoyl) -2- (2-naphthyl) ethyl) -N-methyl amide

(2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (4-iodophenyl)

Can J. Chem. (1977), < / RTI > 55,906.

¹ H-NMR: (CDCl ₃ ) d 1.34 (s, 4.5H), 1.38 (s, 4.5H), 2.70 (s, 1.5H), 2.75 (s, 1.5H); 2.85-3.10 (m, 1 H), 3.2-3.4 (m, 1 H); (M, 2H), 7.62 (d, J = 10 Hz, 2 Hz), 9.5-10 (bs,

N - ((1 R) -2- (4-iodophenyl) -1- (methylcarbamoyl) ethyl-N-methylcarbamic acid tert- butyl ester

(2R) -2- (N-tert-butoxycarbonyl-N-methylamino) -3- (4-iodophenyl) propionic acid (2.00 g, 4.9 mmol) was dissolved in methylene chloride (20 ml). Hydroxybenzotriazole hydrate (0.67 g, 4.9 mmol) and 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (0.99 g, 4.9 mmol) were added and the mixture was stirred for 15 min. Methylamine (0.38 g of a 40% solution in methanol, 4.9 mmol) was added and the mixture was stirred overnight. Methylene chloride (40 ml) was added and the mixture was washed with a saturated aqueous solution of sodium hydrogencarbonate (50 ml) and a sodium hydrogen sulfate solution (10%, 50 ml). The organic phase was dried (MgSO ₄₎ and the solvent was removed in vacuo. The residue was chromatographed on silica (2.5 x 20 cm) using ethyl acetate / heptane (2: 1) as the eluent to give N - ((1 R) -2- (4-iodophenyl) Benzoyl) ethyl) -N-methylcarbamic acid tert-butyl ester (1.77 g).

¹ H-NMR: (CDCl ₃ ) (peak selected for the major rotamer) d 1.39 (s, 9H); 2.75 (s, 3 H); 2.80 (d, 3H); 3.29 (dd, 1 H); 4.88 (t, 1 H).

(2R) -3- (4-iodophenyl) -N-methyl-2- (methylamino) propionamide

N-methylcarbamic acid tert-butyl ester (1.7 g, 4.0 mmol) was dissolved in methylene chloride (10 ml) And trifluoroacetic acid (5 ml) was added. The mixture was stirred for 1 h. Methylene chloride (30 ml) and water (30 ml) were added. Solid sodium bicarbonate was added to pH 8. The organic phase was separated, dried (MgSO ₄ ) and evaporated in vacuo to give 1.22 g of (2R) -3- (4-iodophenyl) -N-methyl-2- (methylamino) propionamide.

1 H-NMR: (CDCl ₃ )? 2.28 (s, 3H); 2.68 (dd, 1 H); 2.81 (d, 3H); 3.08-3.19 (m, 2H); 6.95 (d, 2H); 7.63 (d, 2H).

((1R) -1- (methylcarbamoyl) -2- (4-iodophenyl) ethylcarbamoyl) - 2- (2-naphthyl) ethyl) carbamic acid tert-butyl ester

(1.10 g; 3.30 mmol) was dissolved in methylene chloride (10 ml) and HOAt (0.45 g, 3.30 mmol) was added dropwise to a solution of (2R) -2- (N-tert-butoxycarbonyl- g, 3.1 mmol) and EDAC (0.66 g, 3.5 mmol). After stirring for 15 min, a solution of (2R) -3- (4-iodophenyl) -N-methyl-2- (methylamino) propionamide (1.0 g, 3.1 mmol) and diisopropylethylamine ) Was added and the mixture was stirred overnight. Methylene chloride (30 ml) was added and the mixture was washed with a saturated aqueous solution of sodium hydrogencarbonate (30 ml) and a sodium hydrogen sulfate solution (10%, 30 ml). The organic phase was dried (MgSO ₄₎ and the solvent was removed in vacuo. The residue was chromatographed over silica (2.5 x 20 cm) using ethyl acetate / heptane (2: 1) as the eluent to give N-methyl-N - ((1 R) -1- (N- Ethyl) carbamoyl) -2- (2-naphthyl) ethyl) carbamic acid tert-butyl ester as a colorless oil.

¹ H-NMR: (CDCl ₃ ) (peak selected for the major rotamer) d 1.38 (s, 9H); 2.18 (d, 3H); 2.45 (s, 3 H); 2.75 (s, 3 H); 2.75 (s, 3 H); 5.05 (m, 1 H); 5.42 (m, 1 H).

(2R) -N- (1R) -2- (4-iodophenyl) -1- (methylcarbamoyl) ethyl) -N- amides

((1R) -1- (methylcarbamoyl) -2- (4-iodophenyl) ethylcarbamoyl) - 2- (2-naphthyl) ethyl) carbamic acid tert-butyl ester was dissolved in a mixture of methylene chloride and trifluoroacetic acid and stirred for 15 minutes.

Methylene chloride (20 ml) and water (30 ml) were added. Solid sodium bicarbonate was added to pH 8. The organic phase was separated, dried (MgSO ₄₎ and evaporated in vacuo (2R) -N - ((1R ) -2- (4- iodophenyl) -1- (methylcarbamoyl) ethyl) -N- methyl- To obtain 1.40 g of 2-methylamino-3- (2-naphthyl) propionamide.

¹ H-NMR: (CDCl ₃ ) (selected peak for major rotamer) d 1.79 (s, 3H); 2.02 (d, 3H); 2.55 (s, 3 H); 3.78 (dd, 1 H); 5.44 (dd, 1 H).

(4-iodophenyl) -1- (methylcarbamoyl) ethyl) -N-methylcarbamoyl) -2 - (2-naphthyl) ethyl) -N-methylcarbamoyl) piperidine-1-carboxylic acid tert-butyl ester

1-tert-Butoxycarbonylpiperidine-4-carboxylic acid (143 mg, 0.66 mmol) was dissolved in methylene chloride (10 ml) and HOAt . After stirring for 15 minutes, a solution of (2R) -N - ((1 R) -2- (4-iodophenyl) -1- (methylcarbamoyl) -Naphthyl) propionamide (350 mg, 0.66 mmol) and diisopropylethylamine (85 mg, 0.66 mmol) were added and the mixture was stirred overnight. Methylene chloride (20 ml) was added and the mixture was washed with a saturated aqueous solution of sodium hydrogencarbonate (20 ml) and a sodium hydrogen sulfate solution (10%, 20 ml). The organic phase was dried (MgSO ₄₎ and the solvent removed in jingo. The residue was chromatographed on silica (2.5 x 20 cm) using ethyl acetate as the eluant to give 4- (N - ((1 R) -1- (N - ((1 R) -2- (4-iodophenyl) (2-naphthyl) ethyl) -N-methylcarbamoyl) piperidine-1-carboxylic acid tert-butyl ester 412 mg.

(4-iodophenyl) -1- (methylcarbamoyl) ethyl) -N-methylcarbamoyl) -2 Carboxylic acid tert-butyl ester (412 mg, 0.56 mmol) was dissolved in methylene chloride (5 ml) and trifluoroacetic acid (5 ml), to which was added a solution of 2- Dissolved in the mixture and stirred for 5 minutes. Methylene chloride (20 ml) and a saturated aqueous solution of sodium hydrogencarbonate (20 ml) were added. Solid sodium bicarbonate was added to pH 8. Phase was separated The organic phase was dried (MgSO ₄₎ and evaporated to give the title compound 255mg.

¹ H-NMR: (CDCl ₃ ) (selected peak of major rotamer) d 2.32 (d, 3H); 2.58 (s, 3 H); 2.68 (s, 3 H); 5.33 (m, 1 H); 5.84 (t, 1 H).

HPLC: r _t = 33.35 bun (A1)

PDMS: m / z 640.8 (M + H) < ⁺ & gt ^; .

Abbreviation

HBTU O- (benzotriazol-1-yl) -1,1,3,3, -tetramethyluronium hexafluorophosphate

NMP N-methylpyrrolidone

HATU O- (7-azabenzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate

Trt-trityl

HOBT 7-Hydroxybenzotriazole hydrate

3-AmB 3-aminomethylbenzoyl

N-Me-3-AMB 3-methylaminomethylbenzoyl

Claims (20)

Or a pharmaceutically acceptable salt thereof. (I) ABCD (-E) _p Wherein p is 0 or 1; A is hydrogen or R ¹ - (CH ₂ ) _q - (X) _r - (CH ₂ ) _s --CO--, q is 0 or an integer selected from the group: 1, 2, 3, 4, 5; r is 0 or 1; s is 0 or an integer selected from the group: 1, 2, 3, 4, 5; R^OneIs selected from the group consisting of hydrogen, imidazolyl, guanidino, piperazino, morpholino, piperidino or N (R²) -R³, Wherein each R²And R³Is independently hydrogen or lower alkyl optionally substituted with one or more hydroxyl, pyridinyl or furanyl groups; And when r is 1, X is _{-NH-, -CH 2 -, -CH =} CH-, -C (R 16) (R 17) -, , Wherein each R¹⁶And R¹⁷Is independently hydrogen or lower alkyl; B is (G) _t - (H) _u , wherein each of t and u is independently 0 or 1; G, and H are natural L-amino acids or their corresponding D-isomers, or derivatives thereof such as 1,4-diaminobutyric acid, amino-isobutyric acid, 1,3-diaminopropionic acid, 4-aminophenylalanine, , 2,3,4-tetrahydroisoquinoline-3-carboxylic acid, 1,2,3,4-tetrahydrounharmane-3-carboxylic acid, N-methyl anthranilic acid, anthranilic acid, N- , Non-natural amino acids such as 3-aminomethylbenzoic acid, 3-amino-3-methylbutanoic acid, sarcosine, nipecotic acid or iso-nipecotic acid; And when t and u are both 1, the amide bond between G and H is optionally replaced by Y-NR ¹⁸ -, wherein Y is -CO- or -CH ₂ - and R ¹⁸ is hydrogen, Aralkyl; C is the formula _{-NH-CH ((CH 2)} W -R 4) and D- amino acids of -CO-, where w is 0, 1 or 2; R ⁴ is Each of which is optionally substituted with halogen, lower alkyl, lower alkoxy, lower alkylamino, amino or hydroxy; D is a D-amino acid of the formula -NR ²⁰ -CH ((CH ₂ ) _k -R ⁵ ) -CO- when p is 1, or D is -NR ²⁰ -CH ((CH ₂ ) and _l -R ⁵⁾ -CH ₂ -R ⁶ or _{^{-NR 20 -CH ((CH 2)}} m -R 5) -CR 6, wherein, k is 0, 1 or 2; l is 0, 1 or 2; m is 0, 1 or 2; R < ²⁰ > is selected from the group consisting of lower alkyl or lower aralkyl; R ⁵ is Each of which is optionally substituted with halogen, lower alkyl, lower alkyloxy, amino or hydroxy; And R ⁶ is piperazino, morpholino, piperidino, -OH or -N (R ⁷ ) -R ⁸ , wherein each R ⁷ and R ⁸ is independently hydrogen or lower alkyl; E is when p is ^{1, -NH-CH (R 10)} - , and (CH _{2) v} -R ^9, wherein v is 0 or an integer selected from the group: 1, 2, 3, 4, 5, 6, 7, 8; R ⁹ is selected from the group consisting of hydrogen, imidazolyl, guanidino, piperazino, morpholino, piperidino, -N (R ¹¹ ) -R ¹² , , Wherein n is 0, 1 or 2 and R < ¹⁹ > is hydrogen or lower alkyl, , Wherein o is an integer selected from the group: 1,2,3, Each R < ¹¹ > and R < ¹² > are independently hydrogen or lower alkyl, or , Each of which is optionally substituted with an optionally halogenated, lower alkyl, lower alkyloxy, amino, alkylamino, hydroxy, or amino group and possibly from a hexapyranos or hexapyranosyl-hexapyranos R¹⁰When p is 1, -H, -COOH, -CH₂-R¹³, -CO-R¹³or -CH₂-OH, < / RTI > wherein < RTI ID = R ¹³ is piperazino, morpholino, piperidino, -OH or -N (R ¹⁴ ) -R ¹⁵ wherein each R ¹⁴ and R ¹⁵ is independently hydrogen or lower alkyl; All amide bonds in the formula (I) except for the bond between C and D are independently selected from -Y-NR ¹⁸ - and can be substituted with, where Y is -CO- or -CH ₂ - and R ¹⁸ is hydrogen, lower alkyl or Lower aralkyl; The following compounds are excluded. (3-Aminomethyl benzoyl) -D-2Nal-N-Me -D-Phe-Lys-NH 2, H-Aib-His-D- 2Nal-N-Me-D-Phe-Lys-NH 2, H-Aib-His-N- Me-D-2Nal-N-Me-D-Phe-Lys-NH 2, 3- (H-Aib-His-D-2Nal-N-Me-D-Phe-NH) N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ((3R) - piperidine-carbonyl) -N-Me-D-2Nal -N-Me-D-Phe-Lys-NH 2, (3-aminomethylbenzoyl) -D-2Nal-N-Me-D-Phe-NH) N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ethane, N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ethane, N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ethane, 3- (H-Aib-His-N-Me-D-2Nal-N-Me- N-Me-D-Phe-NH) -1-morpholinopropane, N-Me- (3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D-Phe- N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ethane, 2 - (((3R) -piperidine carbonyl) -D-2Nal-N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ethane. 2. A compound according to claim 1, wherein A is hydrogen, 3-AMB, N-Me-3-AMB or Aib. 2. A compound according to claim 1 wherein t is 1 and u is 0 and G is selected from the group consisting of 3-aminomethylbenzoyl, nifecotic acid and isonipecotic acid. 2. The compound according to claim 1, wherein t is 1, u is 1, G is Aib and H is selected from the group consisting of His, Phe and Ala. 2. A compound according to claim 1, wherein C is selected from the group consisting of D-2-Nal and D-Phe. 2. Compounds according to claim 1, wherein when p is 1, D is D-Phe or D-2Nal. According to claim 1, p is 0, D is the compound wherein D-Phe-NH ₂ or the D-2Nal-NH _2. According to claim 1, E is when p is ^{1, -NH-CH (R 10)} - , and (CH _{2) v} -R ^9, wherein v is 0 or the group: is selected from the integers 1, 2, 3, 4 ego; R ⁹ is hydrogen, morpholino, piperidino, N (R ¹¹ ) -R ¹² , Wherein n is 0, 1 or 2 and R ¹⁹ is hydrogen or lower alkyl, each R ¹¹ and R ¹² is independently hydrogen or lower alkyl, and R ¹⁰ is selected from the group consisting of -H, -COOH, -CH ₂ -R ¹³ , -CO-R ¹³ or -CH ₂ -OH when p is 1, wherein R ¹³ is piperazino, morpholino, piperidino, -OH or -N (R ¹⁴⁾ -R ^15, wherein each of R ¹⁴ and R ¹⁵ are independently a compound, characterized in that hydrogen or lower alkyl. The method of claim 1, wherein at least one of the amide bonds between A and B, between B and C, between D and (E) p, and between G and H is substituted with -CO-N (CH ₃ ) - . Or a pharmaceutically acceptable salt thereof. (R) -2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me) -3-phenylpropanol or its TFA salt; 3 - ((3-aminomethylbenzoyl)) N-Me-D-2Nal-N-Me-D-Phe-NH) -1-1N, N-dimethylaminopropane or a TFA salt thereof; N-Me-D-Phe-NH) -1-N, N-dimethylaminopropane, or its TFA salt; N-Me-D-Phe-NH) - (1-methyl-2-pyrrolidinyl) ethane, or Its TFA salt; H-Aib-His-D- 2Nal-N-Me-D-Phe-Ser-NH 2, or its TFA salt; (3-Aminomethyl benzoyl) -D-2Nal-N-Me -D-Phe-Lys-NH 2, or its TFA salt; (4-piperidine-carbonyl) -D-2Nal-N-Me -D-Phe-NH 2, or its TFA salt; ((3R) -3- piperidin-carbonyl) -D-2Nal-N-Me -D-Phe-NH 2, or its TFA salt; (3-Aminomethyl benzoyl) -D-Phe-N-Me -D-Phe-NH 2, or its TFA salt; (3-aminomethyl-benzoyl) -N-Me-D-Phe -N-Me-D-Phe-Lys-NH 2, or its TFA salt; ((3R) -3- piperidin-carbonyl) -N-Me-D-Phe -N-Me-D-Phe-Lys-NH 2, or its TFA salt; H-Aib-His-N- Me-D-Phe-N-Me-D-Phe-Lys-NH 2, or its TFA salt; ((3R) -3- piperidin-carbonyl) -N-Me-D-2Nal -N-Me-D-Phe-NH 2, or its TFA salt; (2R) -2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me) -3- (2-naphthyl) propanol or its TFA salt; (3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-NH 2, or its TFA salt; 3 - ((3-aminomethylbenzoyl) -N-Me-D-Phe-NH) -1-N, N- H-Aib-His-N- Me-D-2Nal-N-Me-D-Phe-NH 2, or its TFA salt; (3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-Lys-NH 2, H-Aib-Ala-D- 2Nal-N-Me-D-Phe-Lys-NH 2, or its TFA salt; H-Aib-His-D- 2Nal-N-Me-D-Phe-NH 2, or its TFA salt; 2 - ((3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D- (3-aminomethylbenzoyl) -N-Me-D-2Nal-N-Me-D- N-Me-D-Phe-NH) -1-N, N-dimethylaminopropane, (3-aminomethyl-benzoyl) -N-Me-D-2Nal -N-Me-D-Phe-N-Me 2, H-Aib-His-N- Me-D-2Nal-N-Me-D-Phe-NH 2, 3-aminomethyl-benzoyl -N-Me-D-2Nal- N-Me-D-Phe-NH-CH 3 or its TFA salt; 3-methyl-amino-methyl-benzoyl -N-Me-D-2Nal- N-Me-D-Phe-NH-CH 3, or its TFA salt; H-Aib-His-N-Me-D-2Nal-N-Me-D-Phe-NHMe, Carboxylic acid-N - ((1 R) -1- (N - ((1 R) -2- (4-iodophenyl) -1- (methylcarbamoyl) ethyl) -N -Methylcarbamoyl) -2- (2-naphthyl) ethyl) -N-methyl amide. 10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier or diluent. 12. The composition according to claim 11, wherein the composition is in the form of a unit dose comprising from about 10 to about 200 mg of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof. A pharmaceutical composition for stimulating the release of growth hormone from the pituitary gland, comprising a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier or diluent ≪ / RTI > 14. A pharmaceutical composition according to claim 11, 12 or 13 for oral, transdermal, nasal, pulmonary or parenteral administration. A method for stimulating the release of growth hormone from the pituitary gland, comprising administering to the subject a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to any one of claims 11, 12 or 14 Comprising administering to a patient in need thereof an effective amount of a composition according to claim < RTI ID = 0.0 > 1. < / RTI > 16. A method according to claim 15, wherein the effective amount of a compound according to any of claims 1 to 10, or a pharmaceutically acceptable salt or ester thereof is from about 0.0001 to about 100 mg / kg body weight per day, preferably from about 0.001 to about 50 mg / kg body weight. A method for increasing the rate and degree of growth of an animal, increasing milk or parental production of an animal, or treating a disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 10, Or a pharmaceutically acceptable salt thereof, or an effective amount of a composition according to any one of claims 11, 12 or 14 to a patient in need thereof. 10. A compound according to any one of claims 1 to 10 for use as a medicament, or a pharmaceutically acceptable salt thereof. Use of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for stimulating the release of growth hormone from the pituitary gland. A compound according to any one of claims 1 to 10 for the manufacture of a medicament for increasing an animal's growth rate and degree, for increasing milk or parental production of an animal, or for administering to an animal for the treatment of a disease Or a pharmaceutically acceptable salt thereof.