HUT75224A - Combination of bisphosphonates and growth hormone secretagogues - Google Patents

Description

Xl ^ COMBINATIONS OF PHOSPHONATES AND GROWTH Hormone Selection Agents

Bisphosphonates (bisphosphonic acids) are known to inhibit bone resorption and can be used to treat bone disease (U.S. Pat. No. 4,621,077 to Rosini et al.). The treatment of osteoporosis with calcitonin alone or in combination with human growth hormone has been investigated by Aloia et al., 1985, Metabolism 34 (2) 124-129. No improvement in the treatment of osteoporosis has been attributed to the combination of calcitonin therapy with growth hormone dosing in this publication. The effect of growth hormone itself in the treatment of osteoporosis was investigated by Aloia et al. Clin. Endocrinol. Metab., 54, 992-999 (1976)]. Certain non-peptide growth hormone secretagogues are known to stimulate the pituitary gland to increase growth hormone secretion in children and adults with growth hormone deficiency, severe burn injuries, reversal of the adverse effects of glucocorticoid treatment, and growth hormone deficiency. used to treat muscle and exercise deficits, and to treat osteoporosis. Growth hormone secretion

83592-2037 Ski

- compounds having 2 enhancing activities are described, for example, in U.S. Patent Nos. 3,239,345, 4,040,979, 4,418,1890, 5206235, 5,282,441, 5,310,737, 5,317,017; EP 0144230, EP 0513974, WO 94/07486, WO 94/08583, WO 94/13696 and Science 260, 1640-1643 (1993). Other compounds having growth hormone secretagogue activity are described herein.

Numerous bisphosphonic acid derivatives are known in the art for use in the treatment and prevention of bone resorption disorders. Examples of these can be found in US 3251907, US 3422137, US 3584125, US 3940436, US 3944599, US 3962432, US 4054598, US 4267108, US 4327039, US 4407761, US 4578376, US 4621077, US 4624947, US 4746654, U.S. Patent No. 4,761,406; U.S. Pat. No. 4,002,2007; U.S. Pat. No. 4,942,157; U.S. Pat.

EP 0252504 and J. Org. Chem., 36, 3843 (1971).

The preparation of bisphosphonic acids and halogen-bisphosphonic acids is well known in the art. By way of example, mention may be made of the aforementioned literature disclosing the use of the above compounds for the treatment of disorders of calcium or phosphate metabolism, in particular as an inhibitor of bone resorption.

The present invention relates to combinations of bisphosphonate (bisphosphonic acid) and growth hormone secretagogues for the treatment or prevention of disorders of calcium and phosphate metabolism, in particular for the treatment and prevention of bone resorption disorders, in particular osteoporosis, Paget's disease, Paget's disease. and for the treatment and prevention of metastatic bone disease. This particular combination produces unexpected results in the treatment and prevention of the above clinical disorders. Furthermore, it was an object of the invention to identify within the above types the preferred compounds which can be used in the combinations of the invention. It was a further object of the present invention to provide compositions containing the above compounds for use in the treatment of osteoporosis. Other objects of the invention will become apparent from the following description.

The bisphosphonate compound, in particular a bisphosphonate (X) of the formula (X), is one of the components of the combination according to the invention for the treatment and prevention of osteoporotic diseases, in particular osteoporosis in the elderly.

R ¹ is

(a) C 1-5 alkyl, unsubstituted or substituted with one of the following:

1) nh ₂ ,

2) pyridyl,

3) pyrrodinyl,

4) NR ³ R ⁴

(b) NR ⁵ ,

(c) SR ⁶ or

d) Cl;

R ² is H, OH or Cl;

R ³ is H or C 1-4 alkyl,

- 4 R ⁴ is C 1 -C 4 alkyl,

R ⁵ is C 1 -C 10 alkyl and

^R6 contains an aryl group, or a pharmaceutically acceptable salt thereof.

Compounds of formula (X) also include salts of the above compounds with alkali metals, organic bases and basic amino acids.

Of the above compounds of formula (X), those in which

R ¹ is a C 1 -C 5 alkyl group most preferably substituted on its terminal carbon by an amino group or a pyridyl group, and

R ² is hydroxy.

Salts with alkali metals are preferred, and salts with sodium are most preferred.

The most preferred bisphosphonates are selected from the group consisting of alendronic acid, etidronic acid, clodronic acid, pamidronic acid, tiludronic acid, rizedronic acid, 6-amino-1-hydroxyhexylidene bisphosphonic acid and 1-hydroxy-3- (methylpentylamino) bisphenylpropyl and pharmaceutically acceptable salts thereof.

The preparation of bisphosphonic acids is well known in the art. By way of example, reference is made to US 3251907, US 3422137, US 3584125, US 3940436, US 3944599, U.S. Pat.

3962432, US 4054598, US 4267108, US 4327039, US 4407761, US 4578376, US 4621077, US 4624947, US 4746654, US

U.S. Patent Nos. 4,761,406, 4,922,797, 4,942,157, 5,227,506; EP 0252504 and J. Org. Chem., 36, 3843 (1971).

The second element of the combination for treating osteoporosis according to the invention is a growth hormone secretagogue.

Such growth hormone secretagogues are described, for example, in U.S. Patent Nos. 3,239,345, 4,040,979, 4,411,890, 5206235, 5284841, 5,310,737, 5,317,017; EP 0144230, EP 0513974, WO 94/07486, WO 94/08583, WO 94/13696 and Science 260, 1640-1643 (1993).

U.S. Patent 5,202,635 discloses growth hormone secretagogues having the formula: wherein the substituents are as defined in the above cited patent.

The most preferred benzolactam compounds disclosed herein are compounds of formula XII or XIII.

Further growth hormone secretagogues are disclosed in WO 94/13696, which are spiro compounds of formula I or II, in which:

R 1 is C 1 -C 10 alkyl, aryl, (C 1 -C 6) alkylaryl, (C 1 -C 6) alkyl (C 3 -C 7) cycloalkyl, (C 1 -C 5) alkyl-K - (C 1 -C 5) alkyl, (C 1 -C 5) alkyl-K (C 0 -C 5) alkylaryl, (C 1 -C 5) alkyl-K (C 5 -C 5) alkyl - (C 3 -C 7) cycloalkyl, wherein

- 6 K represents Ο, S (O) _m , N (R ₂ ) C (O), C (O) N (R ₂ ), OC (O), C (O) O or -CR ₂ = CR ₂ - or -OC- and aryl are as defined below, and R ₂ and alkyl are further substituents

1-9 halogens, S (O) _{n 1} R _2a , 1-3 OR _2a or C (O) OR _2a , and the aryl groups are further substituted with phenyl, phenoxy, halophenyl, 1-3 6 carbon atoms, 1 to 3 halogen, 1 to 2 _-OR2 group, a methylenedioxy group, -S (O) _m R ₂ groups, 1-2 _CF3 groups, -OCF _3, -NO _2, - N (R ₂ ) (R ₂ ),

-N (R ₂ ) C (O) R ₂ , -C (O) OR ₂ , -C (O) N (R ₂ ) (R ₂ ), -SO ₂ N (R ₂ ) (R ₂ ), - They may contain N (R ₂ ) S (O) ₂ -aryl and -N (R ₂ ) SO ₂ R ₂ ;

R ₂ is hydrogen, C 1-6 alkyl,

C 3 -C 7 cycloalkyl, and where two C 1 -C 6 alkyl groups are present on an atom, they may optionally form a C 3 -C 8 ring, which may also contain oxygen, sulfur or NR _2a ;

R _2a is hydrogen or C 1-6 alkyl;

R _3a and R _3b are independently hydrogen, halogen, C 1 -C 6 alkyl, -OR ₂ , cyano, -OCF ₃ , methylenedioxy, nitro, -S (O) _m R, -CF, or -C (O) OR ₂ , and

- when R _3a and R _{3b are in an} ortho-flux, they may together form a C 5 -C 8 aliphatic or aromatic ring optionally containing 1 or 2 heteroatoms selected from oxygen, sulfur and nitrogen;

R ₄ and R ₅ are each independently hydrogen, C 1-6 alkyl, C 1-6 alkyl having 1-5 halogens, 1-3 hydroxy,

C 1-3 (C 1 -C 10) alkanoyloxy, C 1-3 (C 1 -C 6) alkoxy, phenyl, phenoxy, 2-furyl, (C 1-6) alkoxycarbonyl, (1-6) C 1) alkyl substituted with -O _m -S; obsession

R ₄ and R _{5 taken} together may form - (CH ₂ ), La (CH ₂ ) _s -, wherein

L _a is -C (R ₂ ) ₂ -, -O-, -S (O) _m - or -N (R ₂ ) -, wherein r and s are independently 1-3 and R ₂ is as defined above;

R ₆ is hydrogen or C 1-6 alkyl;

A is a group of formula (a) or (b) wherein x and y are independently 0-3,

R ₇ and R _7a are each independently hydrogen, C 1-6 alkyl, -OR ₂ , trifluoromethyl, phenyl, substituted C 1-6 alkyl, wherein the substituents are imidazolyl, phenyl, indolyl, p-hydroxy-phenyl; 8 nyl, -OR ₂ , 1-3 fluoro,

-S (O) _m R ₂ , -C (O) OR ₂ , C 3 -C 7 cycloalkyl, -N (R ₂ ) (R ₂ ), -C (O) N (R ₂ ) (R ₂ ) , obsession

R ₇ and R _7a independently of one another may be linked to one or both of R ₄ and R ₅ to form an alkylene bridge between the terminal nitrogen atom and the alkyl moiety of R ₇ and R _7a which may contain from 1 to 5 carbon atoms;

B, D, E and F are independently -C (R ₈ ) (R ₁₀ ) -, -O-,

C = O, -S (O) _m - or -NR ₉ - such that one or two of B, D, E and F may be absent to give a 5-, 6- or 7-membered ring ; and with the proviso that B, D, E and F can only be -C (R ₈ ) (R ₁₀ ) - or C = O if one of the remaining groups B, D, E and F is -0- , -S (O) _m - or -NR ₉ -, or

B and D, or D and E together may be -N = CR ₁₀ - or -CR ₁₀ = N-, or B and D, or D and E together may be -CR ₈ = CR ₁₀ ; with the proviso that one of the other B and E or F is at the same time -O-, -S (O) _m - or -NR 1 -;

R ₈ and R ₁₀ are independently hydrogen, -R ₂ , -OR ₂ , - (CH ₂ ) _q -aryl, - (CH ₂ ) _q -C (O) OR ₂ , - (CH ₂ ) _q -C ( O) O (CH ₂ ) _q -aryl or - (CH ₂ ) _q - (1H-tetrazol-5-yl) wherein the aryl group has 1 to 3 halogens, 1 or 2 C 1 -C 8 alkyl groups, 1 to 3 OR ₂ or 1 or 2 -C (O) O R ₂ groups;

- 9 R 1 represents -R ₂ - (CH ₂ ) _q -aryl, -C (O) R ₂ , -C (O) (CH ₂ ) _q -aryl,

-SO ₂ R ₂ , -SO ₂ (CH ₂ ) _q -aryl, -C (O) N (R ₂ ) (R ₂ ), -C (O) N (R ₂ ) (CH ₂ ) _q -aryl, -C (O) OR ₂ , 1 -H-tetrazol-5-yl, -SO ₃ H, -SO ₂ NHC 1 -N, -SO ₂ N (R ₂ ) -aryl, -SO ₂ N (R ₂₎ ) (R ₂ ) and the moiety (CH ₂ ) _q may be optionally substituted with 1 or 2 C 1 -C 4 alkyl groups, and R ₂ and aryl may be optionally substituted with one or more substituents selected from the group consisting of: OR _2a , -O (CH ₂ ) _q -aryl, 1-2 -C (O) OR _2a , 1-2 -C (O) O (CH ₂ ) _q -aryl, 1-2 -C (O) N (R _2a ) (R _2a ), 1-2

-C (O) N (R _2a ) (CH ₂ ) _q -aryl, 1-5 halogen, 1-3 C 1-4 alkyl, 1,2,4-triazolyl, 1-H-tetrazole-5- yl, -C (O) NHSO ₂ R _2a , -S (O) _m R _2a , -C (O) NHSO ₂ (CH ₂ ) _q -aryl, -SO ₂ NHC = N, -SO ₂ NHC ( O) R _2a , -SO ₂ NHC (O) (CH ₂ ) _q -aryl, -N (R ₂ ) C (O) N (R _2a ) (R _2a ), -N (R _2a ) C (O) N (R _2a ) (CH ₂ ) _q -aryl, -N (R _2a ) (R _2a ), -N (R _2a ) C (O) R _2a , -N (R _2a ) C (O) (CH ₂₎ ) _q -aryl, -OC (O) N (R _2a ) (R _2a ), -OC (O) N (R _2a ) (CH ₂ ) _q -aryl, -SO ₂ (CH ₂ ) _q CONH- (CH ₂ ) _w NHC (O) R ₁₁₅ wherein w is 2-6 and

_Tl R represents biotin, aryl, or 1 or 2 OR ₂ groups, 1-2 halogen, azido or nitro szubsztitutált aryl;

m is 0, 1 or 2;

n is 1 or 2;

q is optionally 0, 1, 2, 3 or 4; and

G, Η, I, and J are carbon, nitrogen, sulfur, or oxygen, such that at least one of them is hetero10, and one of G, Η, 1, and J may be absent, resulting in a 5- or 6-membered heterocyclic aromatic. a ring is formed;

and pharmaceutically acceptable salts and individual diastereomers thereof.

The terms used in the above structural formulas and throughout the specification have the following meanings.

By alkyl is meant straight-chain or branched carbon groups which may have double or triple bonds. Examples of the above alkyl groups include methyl, ethyl, propyl, ethynyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, isohexyl, allyl, propenyl. , butenyl, butadienyl and the like.

By alkoxy is meant straight or branched chain alkoxy having the specified number of carbon atoms, which may optionally contain double or triple bonds. Examples of the above alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy, allyloxy, propionyloxy. , isobutenyloxy, 2-hexenyloxy and the like.

Halogen means fluorine, chlorine, bromine and iodine.

Aryl is understood to mean aromatic residues of 5- and 6-membered rings containing phenyl, naphthyl and 1-3 heteroatoms, or bicyclic rings formed from fused 5- or 6-membered rings containing 1-3 heteroatoms, the heteroatom being nitrogen, sulfur or sulfur. oxygen. Examples of the above heterocyclic aromatic rings include pyridine, thiophene, benzothiophene, tetrazole, indole, N-methyl-1-indole, dihydroindole, indazole, N-formylindole, benzimidazole, thiazole, furan, pyrimidine and thiadiazole.

Some of the above expressions may occur more than once within a formula, in which case the meanings may be independent of each other.

Most preferred growth hormone secretagogues for use in the combinations of the invention are compounds of formula (V):

R j is a group of the formula (c), (d), (e), (f), (g), (h), (i), (j), (k) or (1);

R _3a is hydrogen or fluorine;

D is -O-, -S-, -S (O) _m -, N (R ₂ ), NSO ₂ (R ₂ ), NSO ₂ (CH ₂ ) - aryl, NC (O) (R ₂ ), NSO ₂ (CH ₂ ) _q OH, NSO ₂ (CH ₂ ) _q COOR ₂ , NSO ₂ (CH ₂ ) _q C (O) -N (R ₂ ) (R ₂ ), N-SO ₂ (CH ₂ ) _q C (O) -N (R ₂ ) (CH ₂ ) _w OH, or (m), (n) or (o), and aryl represents phenyl or pyridyl and the phenyl may be substituted with 1 to 2 halogens. ;

R ₂ is hydrogen or C 1-4 alkyl;

m is 1 or 2;

t is 0, 1 or 2;

q is 1, 2 or 3;

w is 2, 3, 4, 5 or 6;

and the pharmaceutically acceptable salts and certain diastereomeric forms of the above compounds.

The most preferred growth hormone secretagogues for use in the combination of the present invention include:

1) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -2- (1H- indol-3-yl} ethyl) -2-amino-2-methylpropanamide;

2) N- {1 (R) - [(1,2-Dihydro-1-methanecarbonyl-spiro [3H-indole-3,4-piperidin] -1'-yl) carbonyl] -2- (1H-indole) -3-yl} ethyl) -2-amino-2-methylpropanamide;

3) N- {1 (R) - [(1,2-Dihydro-1-benzenesulfonyl-spiro [3H-indol-3,4-piperidin] -1'-yl) carbonyl] -2- (1H- indol-3-yl} ethyl) -2-amino-2-methylpropanamide;

4) N- {1 (R) - [(3,4-Dihydro-spiro [2H-1-benzopyran-2,4'-piperidin] -1'-yl) carbonyl] -2- (1H-indole) 3-yl} ethyl) -2-amino-2-methylpropanamide;

5) N- {1 (R) - [(2-acetyl-1,2,3,4-tetrahydro-spiro [isoquinolin-4,4'-piperidin] -1'-yl) carbonyl] -2- ( indol-3-yl} ethyl) -2-amino-2-methylpropanamide;

6) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyl) oxy) ethyl} -2-amino-2-methylpropanamide;

7) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyl) oxy) ethyl} -2-amino-2-methylpropanamide mesylate salt;

8) N - {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (2 ', 6'-difluoro-phenylmethyl) ethyl} -2-amino-2-methylpropanamide;

- 13 9) Ν- {1 (R) - [(1,2-Dihydro-1-ethanesulfonyl-5-fluoro-spiro [3H-indole-3,4'-piperidin] -1 ') 1) Carbonyl] -2- (phenyl-1-ethyl-1-oxy) -ethyl} -2-amino-2-methylpropanamide;

10) N- {1 (S) - [(1,2-dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyl) thio) ethyl} -2-amino-2-methylpropanamide;

11) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} -2-amino-2-methylpropanamide;

12) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indo-3,4,4'-piperidin] -1'-yl) carbonyl] -3-cyclohexyl -propyl} -2-amino-2-methylpropanamide;

13) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -4-phenyl-butyl} -2-amino-2-methylpropanamide;

14) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3 H -indol-3,4'-piperidin] -1'-yl) carbonyl] -2- (5 -fluoro-1H-indol-3-yl) -ethyl} -2-amino-2-methyl-propanamide;

15) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-5-fluoro-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (5-fluoro-lH-indol-3-yl} ethyl) -2-amino-2-methylpropanamide;

16) N- {1 (R) - [(1,2-Dihydro-1- (2-ethoxycarbonyl) methylsulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl) ] -2- (1H-indol-3-yl) -ethyl} -2-amino-2-methyl-propanamide;

17) N- {1 (R) - [(1,2-Dihydro-1,1-dioxospiro [3H-benzothiophene-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenyl) methyloxy) ethyl} -2-amino-2-methylpropanamide, and pharmaceutically acceptable salts thereof.

A particularly preferred growth hormone secretagogue is, for example

N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyl) oxy) ethyl} -2-amino-2-methylpropanamide mesylate salt; the

N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} - 2-amino-2-methylpropanamide, and pharmaceutically acceptable salts thereof.

The preparation of bisphosphonic acids is well known in the art. Various bisphosphonic acid compounds and methods for their preparation are described, for example, in US 3251907, US 3422137, US 3584125, US 3940436, US 3944599, US 3962432, US 4054598, US 4267108, US 4327039, US 4407761 and US 4621077. U.S. Patent 4,746,654; U.S. Pat. No. 4,624,497; U.S. Pat. EP 0252504 and J. Org. Chem., 36, 3843 (1971).

The preparation of growth hormone secretagogues is described in detail in the following literature:

U.S. Pat. Nos. 3,239,345, 4,036,979, 4,411,890, 5206235, 5,282,441, 5,310,737 and 5,317,017; EP 0144230, EP 0513974, WO 94/07486, WO 94/08583, WO 94/13696 and Science 260, 1640-1643 (1993).

The growth hormone secretagogue of the formulas (I) and (II) used in the combinations of the invention may be prepared by two different synthetic methods. Syntheses for the preparation of compounds of formulas I and II are illustrated in detail by the following reaction schemes.

In most cases, the protected amino acid derivatives of formula (1) are commercially available when the protecting group L is, for example, a BOC or CBZ group. Other protected amino acid derivatives of formula (I) may be prepared by methods known in the art. Most of the spiropiperidines and spiroazepines (n = 2) of formulas (2) and (2a) are known in the art and are conventionally used on the phenyl or heteroaryl group, e.g. derivatized. Alternatively, the various phenyl or heteroaryl-substituted spiropiperidines and spiroazepines (n = 2) may be prepared by methods known in the art using derivatized phenyl and heteroaryl intermediates. In the Schemes following Scheme 1, synthetic procedures are only illustrated with spiropiperidines, although it will be obvious to those skilled in the art that the transformations shown may be carried out with higher homologues to give compounds of Formulas I and II wherein n = 2.

Intermediates (3) and (3a) may be synthesized according to Scheme 1. The coupling of the spiropiperidines of formulas 2 and 2a to the protected amino acids of formula I, wherein L is a suitable protecting group, is conveniently carried out in an inert solvent such as dichloromethane in the presence of a coupling agent such as DCC or EDC in the presence of HOBT. Alternatively, the coupling may be carried out with a coupling solvent such as BOP in an inert solvent such as dichloromethane. Separation of the unwanted by-products and purification of the intermediates can be carried out by chromatography on silica gel by flash chromatography [W. C. Still, M. Kahn, and A. Mitra, J. Org. Chem. 43, 2923 (1978)] using MPLC or preparative TLC.

The conversion of intermediates (3) and (3a) into intermediates (4) and (4a) can be carried out according to Scheme 2. Removal of benzyloxycarbonyl groups can be accomplished by a number of known methods; for example by catalytic hydrogenation with hydrogen in the presence of a palladium or platinum catalyst in a protic solvent such as methanol. If catalytic hydrogenation is not recommended due to the presence of other potentially reactive functional groups, the removal of the benzyloxycarbonyl groups may be accomplished by treatment with hydrobromic acid in acetic acid. Removal of the BOC protecting groups can be accomplished in a solvent such as methylene chloride or methanol with a strong acid such as hydrochloric acid or trifluoroacetic acid. Known reaction conditions may also be used to remove any other protecting groups that may be present. Green T., Wuts P.G.M. Protective Groups in Organic Synthesis, John Wiley & Sons, Inc., New York, NY (1991)].

Intermediates (5) and (5b), wherein A is methylene or substituted methylene, can be prepared according to Scheme 3 by linking intermediates (4) or (4a) with amino acids (6). , again in an inert solvent such as di17 chloromethane, with a coupling agent such as EDC or DCC in the presence of HOBT. The amino acids of formula (6) are either known or can be readily prepared by synthetic methods known in the art. Alternatively, the coupling may also be carried out with a coupling agent such as BOP in an inert solvent such as dichloromethane. In the case where R ₄ or R ₅ is hydrogen, the coupling reaction uses the amino acids (7) where L is a protecting group for the preparation of intermediates (5a) and (5c). Deprotection (L = protecting group) of the compounds of formula (5a) and (5c) can be carried out under known reaction conditions.

Compounds of formula (I) and (II) wherein R ₄ and / or R ₅ are hydrogen may be converted to new compounds of formula (I) and (II) (preferred side chain is R ₇ = CH ₂ -CH (OH)). -CH ₂ X wherein X = H or OH] substituted at the amino group according to Scheme 4. Reductive amination of compounds of formula (I) and (II) with an aldehyde can be carried out under known conditions; for example, by catalytic hydrogenation with hydrogen in the presence of a platinum, palladium or nickel catalyst, or with chemical reducing agents such as sodium cyanoborohydride in a solvent such as methanol or ethanol. Alternatively, a similar conversion may be made by an epoxide ring-opening reaction.

(I) and (II) compounds of formula I, wherein _Z is N (R ₂₎ (CH ₂₎ -C _{(R7) (R7a)} - _{(CH2) y,} can be prepared according to Scheme 5, in such a way reacting a compound of formula (4) or (4a) with a reagent of formula (8), wherein

X is an easily removable group such as chloro, bromo, iodo or imidazolyl.

Alternatively, the compounds of formula (4) and (4a) may also be reacted with the isocyanate of formula (9) in an inert solvent such as 1,2-dichloroethane. When R ₄ and R ₅ are hydrogen in the final product, the reagents (8) and (9) contain L removable protecting groups instead of R ₄ or R ₅ .

The compounds of formula (I) and (II) of the present invention may also be prepared in a convergent manner as described in Schemes 6, 7 and 8.

In many cases, the protected amino acid derivatives of formula (10) are commercially available when M is methyl, ethyl or benzyl ester. Other ester protected amino acids may be prepared by known methods. Some of the above procedures involve the reaction of the protected amino acid with a diazoalkane and deprotection of the L, reaction of the amino acid with a suitable alcohol in the presence of a strong acid such as hydrochloric acid or p-toluenesulfonic acid. Synthetic pathways for the preparation of new amino acids are shown in Schemes 14, 15 and 16.

Intermediates (11) and (11a) can be prepared according to Scheme 6 by linking the amines (10) with the amino acids (6) and / or (7) in which L is a protecting group, as described in Scheme 3. reaction19 as outlined in Scheme 19. If a urea bridge is present in the compound of formula (11) or (11a), it may be introduced as described in Scheme 5.

The esters of formula (11) or (11a) can be converted to the acid intermediates of formula (12) or (12a) by a number of known methods according to Scheme 7; For example, methyl and ethyl esters may be hydrolyzed with lithium hydroxide in a protic solvent such as aqueous methanol. In addition, the removal of the benzyl group can be accomplished by various reduction processes, such as hydrogenation in the presence of a platinum or palladium catalyst in a protic solvent such as methanol. The allyl ester may be cleaved by a tetrakis triphenylphosphine palladium catalyst in the presence of 2-ethylhexanoic acid in various solvents such as ethyl acetate and dichloromethane [see, e.g., J. Org. Chem. 42, 587 (1982)].

The acids (12) and (12a) can then be converted to the compounds (5) and (5a) and (5b) and (5c) as described in Scheme 8. The coupling of the spiropiperidines of formula (2) and (2a) with the acids of formula (12) or (12a), wherein L is a suitable protecting group, is conveniently carried out in an inert solvent such as dichloromethane with a coupling agent such as dicyclohexylcarbodiimide (DCC) EDC in the presence of 1-hydroxybenzotriazole (HOBT). Alternatively, the coupling may be carried out with a coupling agent such as benzotriazol-1-yloxytris (di-methylamino) phosphonium hexafluorophosphate (BOP) in an inert solvent such as dichloromethane.

Compounds of formula (5a) and (5c) are converted to compounds of formula (I) and (II) by deprotection of L. When R ₄ and / or R ₅ is hydrogen, optionally substituted alkyl groups on the nitrogen may be introduced.

4.

Preparation of the oxo-containing spiroindanylpiperidine intermediates can be carried out according to Scheme 9 for compounds wherein R _3a and R _3b are hydrogen. The protected spiroindan (13) is hydroboronated and then oxidatively treated with pyridinium chlorochromate to give the spiroindanone (14).

The conversion of spiroindans to benzolactam intermediates is illustrated in Scheme 10. Treatment of spiroindanone with hydrazonic acid in an inert solvent such as chloroform (Schmidt reaction) is one of several known methods for conversion. In this example, a mixture of the two benzolactams is formed. The isomers can be easily separated by chromatography on silica gel, and the resulting intermediates, after deprotection, are incorporated into growth hormone secretagogues according to Schemes 1 and 8, using intermediates of formula (2).

Intermediates (15) and (16) are alkylated with an alkyl halide in a solvent such as dimethylformamide in the presence of sodium hydride to give compounds (17) and (18) (R ₂ = C 1 -C 4 alkyl). The process is illustrated in Scheme 10A.

When L is a suitable protecting group, such as benzyl, reduction of the amides with lithium aluminum hydride 21 affords the amines of formulas 19 and 21. These amines wherein R ₂ is hydrogen may be alkylated, acylated, or reacted with substituted sulfonyl halides or isocyanates under known reaction conditions to provide compounds of formulas 20 and 22 (see Schemes 11 and 11A). The L protecting group is then removed by hydrogenolysis using a palladium catalyst and the resulting intermediates can then be introduced into the growth hormone secretagogues of the present invention as shown in Schemes 1 and 8 using the intermediates of formula (2).

The 1,2,3,4-tetrahydro-spiro [isoquinoline-4,4'-piperidine] ring system may also be prepared according to Scheme 12. Ozonolysis of the protected spiroindene followed by treatment with dimethylsulfide affords the hemiacetal intermediates of formula (24) to give the amine of formula (25) by reductive amination and acylation. The amino protecting group is as defined above.

Ring analogs of the general formula wherein X, Y = H, H;

ΟΗ, Η; H, OH; and = O, may be prepared by methods known in the art. For example, according to the procedure outlined in Scheme 13, the spiro [2H-1-benzopyran-2,4'-piperidine] analog can be prepared from a substituted or unsubstituted 2-hydroxyacetophenone and a suitably protected 4-piperidone.

H.J. (See Synthesis 1978, 886-887 and references cited therein). The 2-hydroxyacetophenones are either commercially available or may be prepared by known methods. Such procedures are described, for example, in Chang C.T. and

- 22 [J. Chem. Soc., 3414-3417 (1961)] and Elliott, J.M., et al., J. Am. Med. Chem. 35: 3973-3976 (1992). Deprotection is also known in the art (Protective Groups in Organic Synthesis, Greene T.W., Wuts P.G., John Wiley & Sons, New York, 1991); and Olofson, R.A., et al., J. Org. Chem. 49, 2081-2082 (1984)] to give the amine which can then be incorporated into the growth hormone secretagogues of the present invention using the intermediates of formula (2) as described in Schemes 1 and 8.

The ketone group of the compound of formula (27) can be reduced to alcohol using sodium borohydride or completely reduced to the methylene group, also under known conditions. For example, reduction of the ketone with sodium borohydride followed by treatment with concentrated hydrochloric acid and hydrogenation affords compounds of formula (29). The amines (27), (28) or (29) can then be incorporated into the growth hormone secretagogues of the present invention using the compounds of Formula (2) following the procedure outlined in Schemes 1 and 8. Alternatively, the ketone may be reduced after conversion to the compounds of formula (I).

Chiral hydroxy-spiro [2H-1-benzopyran-2,4'-piperidine] analogs can be prepared using optically active reducing agents and crystallization of diastereomeric salts.

The compounds of formula (I) and (II) of the present invention may be prepared from various substituted natural and non-natural amino acids, such as those of formulas (30), (6) and (7), wherein A is - (CH ₂ ) _x -. C (R ₇ ) (R _7a ) - 23 - (CH ₂ ) _y -. The preparation of several of the above amino acids is also described in U.S. Patent 5,206,237.

Several methods for preparing the above products in racemic form are known (Williams, R. M., "Synthesis of Optically Active α-Amino Acids", Pergamon Press, Oxford, Vol. 7, 1989). There are several methods for resolving (DL) amino acids of formula (30). According to one of the known processes, the amino- or carboxyl-protected intermediates are resolved by crystallization of salts prepared with optically active acids or amines. Alternatively, the amino group of the carboxyl-protected intermediates may be coupled with optically active acids by the methods described above. Separation of the individual diastereomers is accomplished either by chromatography or by crystallization followed by hydrolysis of the chiral amide to give the resolved amino acids. Similarly, the amino-protected intermediates can be converted into a mixture of chiral diastereomeric esters and amides. Separation of the mixture can be accomplished by the above methods and hydrolysis of each diastereomer gives the (D) and (L) amino acids. Finally, an enzymatic process for the resolution of N-acetyl derivatives of (DL) amino acids is also known (Whitesides et al., 1989, J. Am. Chem. Soc., 11, 6354-6364).

If intermediates are to be prepared in optically pure form, several methods are known, for example

1) asymmetric electrophilic amination of chiral enolates [J. Chem. Soc. 108, 6394-6395, 6395-6397 and 6397-6399 (1986)];

- 24 2) asymmetric nucleophilic amination of optically active carbonyl moieties [J. Am. Chem. Soc., 1992, 114, 1906; Tetrahedron Lett. 28, 32 (1987)];

3) diastereoselective alkylation of chiral glycine enolate fibrous plastics [J. Chem. Soc., 13, 9276 (1991); J. Org. Chem., 54, 3916 (1989)];

4) diastereoselective nucleophilic addition to chiral electrophilic glycinate filaments [J. Chem. Soc. 108, 1103 (1986)];

5) asymmetric hydrogenation of prochiral dehydro-amino acid derivatives (Asymmetric Synthesis, Chiral Catalysis; Morrison, J.D., ed., Akademic Press: Orlando, FL, 1985; vol. 5);

6) enzymatic syntheses [Angew. Chem. Int. Ed. Engl. 1977, 17, 176].

For example, the alkylation of the enolate of diphenyloxazinone (31) with cinnamyl bromide in the presence of sodium bis (trimethylsilyl) amide results in the compound (32) from which the N-tert-butoxycarbonyl group is removal of trifluoroacetic acid and hydrogenation over palladium chloride to give the desired (D) -2-amino-5-phenylpentanoic acid (33) (Scheme 14).

The intermediates of Formula 30, which are O-benzyl (D) -Serine derivatives of Formula (34), are conveniently prepared from appropriately substituted benzyl halides and N-protected (D) -Serine 34. The protecting group L is preferably a BOC or CBZ group. Benzylation of 34 can be accomplished by a variety of methods, such as deprotonation with 2 equivalents of sodium hydride in an inert solvent such as dimethylformamide and treatment with 1 equivalent of benzyl halide (Synthesis 1989, 36) described in Scheme 15. way.

O-alkyl (D) -Serine derivatives are also prepared by alkylation according to Scheme 15. Other methods for the preparation of (D) serine derivatives of formula (35) include acid-catalyzed benzylation of carboxyl-protected intermediates of formula (34) with an ArCH ₂ OC (= NH) CCl ₃ reagent [ SHE. Yonemitsu et al., Chem. Pharm. Bull. 36, 4244 (1988)]. Alternatively, alkylation of chiral glycine enolates [J. Am. Chem. Soc. 113, 9276 (1991); J. Org. Chem. 54, 3916 (1989)] with ArCH ₂ OCH ₂ X where X is a leaving group to give 35. In addition, D, LO-aryl (alkyl) serines can be prepared and resolved by the methods described above.

The alkylation of the N-protected (D) -cysteine of formula (36) can be carried out according to the procedure described for the preparation of (D) -erine derivatives, illustrated in Scheme 16 with an alkylating agent of formula R _1a -X, wherein X is a leaving a group such as halide or mesyloxy.

The oxidation of the cysteine derivative (37) to the sulfoxide (n = 1) (38) or the sulfone (n = 2) (38) can be accomplished by a variety of oxidizing agents. [For oxidation of sulfides see, e.g., Org. Prep. Proc. Int., 14, 45 (1982)]. Sodium periodate [J. Org. Chem. 32, 3191 (1967)] are frequently used for the synthesis of sulfoxides and potassium hydrogen persulfate (OXONE), [Tetrahedron Lett. 22, 1287 (1981)] for the preparation of sulfones.

Again, it is noted that the various substituted amino acids can be incorporated into the growth hormone secretagogues of the present invention by the method outlined in Schemes 1 and 8. Growth hormone secretagogues containing a sulfoxide or sulfone function can also be prepared from cysteine derivative enhancers using sodium periodate or OXONE®. Alternatively, hydrogen peroxide may be used as the oxidizing agent in the final step of the synthesis as shown in Scheme 17. The sulfoxide (n = 1) and sulfone (n = 2) analogs of formula (40) can be separated by preparative thin layer chromatography.

Removal of the amino protecting groups can be accomplished by a number of known methods. See, e.g., Protective Groups in Organic Synthesis, T.W. Greene, John Wiley and Sons, NY (1981)].

Compounds of formula (I) wherein R ₄ and R ₅ are hydrogen may be further reacted as described above by reductive alkylation with aldehydes or by reaction with various epoxides, for example. The products which are obtained in the form of the hydrochloride or trifluoroacetate salt are conveniently purified by reverse phase high performance liquid chromatography (HPLC) or recrystallization.

The spiropiperidines of formula (41) may be prepared by a variety of methods, for example by the syntheses described below.

27 The spiropiperidines of formula (42), wherein L is a protecting group as defined above, may be prepared by known methods (see, e.g., H. Ong et al., J. Med. Chem. 23, 981-986 (1983)). The nitrogen of the indoline 42, wherein L is a protecting group such as methyl or benzyl, may be reacted with various electrophiles to produce the spiropiperidines 43, wherein R _{9 is a} different functionality. Reaction of compounds of formula (42) with, for example, isocyanates yields urea derivatives, reacting with chloroformates in an inert solvent, such as dichloromethane, to obtain carbamates, reacting with acid chlorides, anhydrides or acid imidazoles. reacting with sulfonyl chlorides to give sulfonamides and reacting with sulfamyl chlorides to form sulfamides. In addition, in the compounds of formula (42), the indoline nitrogen may be reductively alkylated with aldehydes under known conditions. When a protected glyoxylic acid of the formula HCOCOOM is used as the aldehyde in the reductive amination reaction, wherein M is a specified protecting group, the M substituent may be removed from the product and further derivatized. Alternatively, compounds of Formula 42 may be reacted with epoxides to provide compounds of Formula 43 wherein R ₉ is β-hydroxy-substituted alkyl or arylalkyl. The indolines of formula 42 can also be converted to the compounds of formula 43 wherein R ₉ is phenyl or substituted phenyl, heteroaryl, or substituted heteroaryl, such that the compound of formula 42 is a fluorophenyl or fluorine. -heteroaryl reagent. These reactions are detailed in H. Ong et al., J. Med.

Chem., 23, 981-986 (1983).

The spiropiperidine intermediate of formula 43 (L = Me or Bn) wherein R ₉ is hydrogen or most of the derivatives described above can be demethylated or debenzylated to give the aforementioned derivative of formula 44 wherein R ₉ is hydrogen, Scheme 19. For compounds of Formula 43 where L = Me, demethylation can be accomplished by a number of known methods. For example, demethylation of compound (43) can be accomplished by reacting the compound with bromocyano and potassium carbonate in an inert solvent such as dichloromethane to reduce the resulting cyanamide to lithium aluminum (44). hydride in tetrahydrofuran, under reflux, or under reflux with a strong acid such as aqueous hydrochloric acid, or with a Grignard reagent such as methyl magnesium bromide. Alternatively, the demethylation of the compound of formula (43) may also be carried out by the ACE-C1 method according to R. Olofson et al., J. Org. Chem. 49, 2795 (1984) and references therein. In the case of intermediates of formula 43 wherein L = Bn, the benzyl group can be removed by reductive methods, for example hydrogenation in the presence of a platinum or palladium catalyst in a protic solvent such as methanol. Alternatively, debenzylation (L = Bn) of compound (43) may also be performed by the ACE-C1 method according to R. Olofson et al., J. Med. Org. Chem (1984).

The spiroheterocyclic compounds of formula (45) may be prepared by a variety of methods, for example, by syntheses according to Scheme 20. The allylic oxidation of the protected piperidine (47) can be accomplished by classical methods [Rabjohn N .: Org. React. 24, 261 (1976)]. The resulting allylic alcohol is treated with thionyl chloride in an inert solvent such as benzene to give the corresponding chloride 48. When D = O or S, the alkylation reaction is carried out using dimethylformamide or acetone as the solvent and potassium carbonate as the base and D = NR ₉ (R ₉ = H, alkyl, aryl, acyl, sulfonyl, carbamate). ) carrying out the reaction with sodium hydride as the base in an inert solvent such as tetrahydrofuran to give the cyclization precursor (49). When L is a designated protecting group, the compounds of formula (49) may be cyclized by a variety of methods. For example, cyclization of the compound of formula (49) may be accomplished by reaction with tributyltin hydride (Curran DP, Synthesis 1988, 417-489) in an inert solvent such as benzene to give the compound of formula (46). Alternatively, the compound of formula 46 (D-NR ₉ ) may be prepared by the procedures outlined in Schemes 18 and 19.

Compounds of formula (46) wherein D = S can be oxidized to a sulfoxide (n = 1) or a sulfone (n = 2) by a variety of oxidizing agents (see Scheme 21). For example, sodium periodate is often used to produce sulfoxides, and sulfones are often used to prepare sulfones.

- 30 OXONE's. Deprotection affords the amines of formula (45) which can then be converted to the growth hormone secretagogues of the present invention using the intermediates of formula (2).

The spiropiperidines (50) and (51) may be prepared according to Scheme 22.

Phthalimides of formula 53 wherein R 1 is alkyl, aryl, - (CH ₂ ) _q -aryl or protecting group are either commercially available or may be prepared from the corresponding phthalimides by methods known in the art (see, e.g., Bewster et al., J. Org. Chem., 28, 501 (1963); Mcalees et al., J. Chem. Soc., 2038 (1977). Phthalimidines of formula 53 may be alkylated in the presence of a base such as potassium hydride, lithium or potassium bis (trimethylsilyl) amide with protected bis (2-haloethyl) amine, where L is a specified protecting group, e.g. methyl, benzyl, t-BOC, or CBZ, etc., and Y is Cl, Br, I, and the reaction product yields the spiropiperidines 54. The protecting group is then deprotected to give compounds of formula (50). Reduction of the lactam ring of the compound (50) with hydrides, such as lithium aluminum hydride, affords the compound (51).

Note that the order in which the reactions described above are performed is not critical, and it will not be difficult for one skilled in the art to select a sequence of reactions that is five ·.

□ ο • «facilitates the reaction or avoids the formation of unwanted by-products.

The use of a compound as a growth hormone secretagogue may be demonstrated by methods known in the art, such as Smith et al., Science 260: 1640-1643 (1993) (see explanation in Figure 2).

The combinations of bisphosphonates and growth hormone secretagogues of the present invention are useful in the therapeutic and prophylactic treatment of disorders of calcium and phosphate metabolism and related disorders. These diseases can be divided into two groups:

1. Abnormal (ectopic) deposition of calcium salts, mainly calcium phosphate, pathological hardening of tissues and bone deformation.

2. Conditions that can be improved by reducing bone resorption. As bone resorption decreases, the balance between absorption and formation should be improved, bone loss should be reduced, or it may result in an increase in bone stock. Decreased bone resorption may relieve pain associated with osteolytic injuries and reduce the incidence and / or magnification of injuries.

The above diseases include: osteoporosis (e.g., estrogen deficiency, immobilization, glucocorticoid-induced or senile), bone formation disorder, Paget's disease, myositis ossificans, Bechterew's disease, malignant hypercalcaemia, metastatic bone disease, metastatic disease, , urinary tract stone disorder, · · · · ···············································································•

- 32 kidney or ureteric or bladder stones, hardening of the arteries (sclerosis), arthritis, bursitis, neuritis and tetany.

Increased bone resorption may lead to abnormally high plasma calcium and phosphate levels, which may also be alleviated by this treatment.

Combination therapy for inhibiting bone resorption, preventing osteoporosis, and promoting healing of fractures can be illustrated by combining bisphosphonates and growth hormone secretagogues according to the present invention. The above applications of bisphosphonates are for example Hamdy N.A.T. Role of Bisphosphonates in Metabolic Bone Diseases, Trends in Endocrinol. Metab., 4, 19-25 (1993)]. Bisphosphonates useful for the above purposes include alendronate, tiludronate, dimethyl APD, rizedronate, etidronate, YM-175, clodronate, pamidronate and BM-210995, the preferred bisphosphonate compound being alendronate.

Combinations of bisphosphonates, particularly pamidronate or alendronate, have an unexpected effect in preventing and treating bone resorption disorders when combined with growth hormone secretagogues. While not wishing to be a particular theory of mode of action, we have shown an enhanced effect in reducing and reversing the rate of bone loss that occurs during aging and is known as osteoporosis with combinations of the above agents that would have been expected individually. In particular, combination therapy with growth hormone secretagogues and bisphosphonates increased bone mass. This is the bone mass that is. Aa aa

33 increase is probably the result of increased bone turnover or bone formation produced by increased growth hormone / lGF-1 concentrations from growth hormone secretagogue and reduced bone resorption by bisphosphonates. According to the literature, such a decoupling of bone formation and bone resorption was not expected.

In a preferred embodiment of the invention, the active substance bisphosphonic acid is 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid (alendronic acid) in combination. Another embodiment of the invention is a dosage form comprising the sodium salt (allendronate) of 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid monosodium salt trihydrate. Our studies show that when administered parenterally, this compound is approximately five times more potent in reducing bone-induced tumor-related hypercalcemia in humans than pamidronate (3-amino-1-hydroxypropylidene-1,1-bisphosphonic acid), which is the most potent available bisphosphonate for the treatment of tumor-induced bone disease. The 4-amino-l-hydroxybutylidene-1,1-bisphosphonic acid and homologues thereof, wherein the R ¹ side-chain is substituted with a variable length from 1 to 5 carbon atoms, N-alkyl, and amino termini can readily be prepared in U.S. 4,407,761 and 4922007. In addition, the mono-, di- and trisodium salts and other pharmaceutically acceptable salts of bisphosphonic acids may also be used in the composition of the present invention, such as ammonium salts, alkali metal salts such as potassium salts, alkaline earth metal salts such as calcium and magnesium salts with amine, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, and the like. skilled salts. These salts may be prepared, for example, by the methods described in U.S. Patent No. 4,922,077.

In the combinations of the invention, the bisphosphonates and growth hormone secretagogues may be administered separately or in combination. In addition, one component may be administered before, during or after the other component.

The components of the combination of the invention may be administered orally, parenterally (e.g., intramuscularly, intraperitoneally, intravenously or subcutaneously by injection or in the form of an implant), nasally, vaginally, rectally, sublingually or topically, and conventional non-toxic, pharmaceutically acceptable or without it, it may be formulated in a suitable dosage unit form.

For administration of the compounds of the present invention, the pharmaceutical compositions are conveniently presented in unit dosage form, which may be prepared by methods well known in the art of pharmacy. All methods include the step of mixing the active ingredients with the carriers and one or more excipients. In general, pharmaceutical compositions are prepared by uniformly and intimately admixing the active ingredients with a liquid carrier or a finely divided solid carrier, or both, and then, if necessary, shaping the product into the desired composition. In the pharmaceutical composition, the active ingredient is present in an amount that is capable of producing the desired effect on the course or symptoms of the disease.

Pharmaceutical formulations suitable for oral administration may contain the active ingredient in physically discrete units, such as hard or soft capsules, tablets, pills, and lozenges, each containing a predetermined amount of the active ingredient; or in the form of dispersible powders or granules; in the form of solutions or suspensions in aqueous liquid or non-aqueous liquid; in the form of syrups or elixirs, or in the form of oil-in-water emulsions or water-in-oil emulsions. Formulations for oral use may be prepared according to conventional techniques for the preparation of pharmaceutical compositions, and may include one or more sweeteners, flavors, colorants, preservatives, to provide a pharmaceutically attractive and acceptable formulation.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is admixed with at least one inert, pharmaceutically acceptable carrier such as sucrose, lactose or starch. The above dosage forms may conventionally contain additional substances other than the inert diluents, for example lubricants such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage form may also comprise buffering agents.

Tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients may also be prepared by known techniques. Suitable excipients include, for example, the following

1) inert diluents, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate;

2) granulating and disintegrating agents such as corn starch and alginic acid;

3) binders such as starch, gelatin or acacia; and

4) lubricants such as magnesium stearate, stearic acid or talc.

The tablets may be uncoated or they may be coated by known techniques to reduce disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, glyceryl monostearate or glyceryl distearate may be used as the sustained release agent. The tablets may also be coated according to the methods described in U.S. Patent Nos. 4,256,108, 4,160,452, and 4,265,874 to form controlled release osmotic pharmaceutical compositions.

In some cases, the compositions for oral administration may also be in the form of hard gelatin capsules containing the active ingredient in admixture with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin. The pharmaceutical compositions of the present invention may also be in the form of soft gelatine capsules in which the active ingredient is mixed with water or an oily medium such as peanut oil, liquid paraffin or olive oil.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents such as water. In addition to the inert diluents mentioned above, these compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring and perfuming agents. Aqueous suspensions generally contain the active ingredient in admixture with excipients suitable for the manufacture of aqueous suspensions. Examples of such excipients include:

1) suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and acacia;

2) dispersing or wetting agents, for example

(a) naturally occurring phosphatides such as lecithin,

(b) condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate,

(c) condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadeca (ethyleneoxy) cetanol;

(d) condensation products of ethylene oxide with partial esters of fatty acids and hexite, such as polyoxyethylene sorbitol monooleate; or

(e) condensation products of ethylene oxide with partial esters of fatty acids and hexite anhydride, such as polyoxyethylene sorbitan monooleate.

Aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

Oily suspensions may be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweeteners and flavors may also be added to provide oral edible preparations. These compositions may also be prepared by the addition of antioxidants such as ascorbic acid.

Dispersible powders and granules are suitable for preparing aqueous suspensions. In these, the active ingredient is mixed with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are, for example, those mentioned above. Additional excipients such as sweeteners, flavoring agents and coloring agents may also be present.

The compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin or a mixture thereof. Suitable emulsifiers include, but are not limited to:

• · · • · · · · · · · · · · · · · · · ·

- 39 (1) naturally occurring rubbers such as acacia and gum tragacanth,

2) naturally occurring phosphatides such as soybeans and lecithin,

3) esters and partial esters of fatty acids and hexite anhydrides, such as sorbitan monooleate,

4) condensation products of the above partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate.

The emulsions may also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol or sucrose. These compositions may also contain plasticizers, preservatives and flavoring or coloring agents.

The pharmaceutical compositions may also be in the form of sterile injectable aqueous or oleaginous suspensions or solutions. The suspensions are prepared by known methods, using suitable dispersing or wetting agents and suspending agents, as mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, in a solution in 1,3-butanediol. Examples of acceptable carriers and diluents are water, Ringer's solution, isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any non-contaminated fixed oil may be employed, for example synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid, may also be used in injectable formulations.

Formulations of the invention suitable for parenteral administration include sterile aqueous and non-aqueous solutions, suspensions, or emulsions. Examples of non-aqueous solvents or vehicles include propylene glycol, polyethylene glycol, vegetable oils such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. Such dosage forms may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. The compositions may be sterilized, for example, by filtration through bacterial retention filters, the incorporation of sterilizing agents into the compositions, or by irradiation of the compositions or by heating the compositions. The compositions may also be prepared in the form of sterile solid compositions which are dissolved in sterile water or other sterile injectable medium immediately prior to use.

The combination of the invention may also be administered rectally in the form of a suppository. This composition can be prepared by mixing the active ingredient with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at rectal temperature and therefore melts in the intestine to release the active ingredient. Examples of such materials are cocoa butter and polyethylene glycols.

Formulations suitable for nasal and sublingual administration are also prepared by conventional techniques.

Combinations of the present invention suitable for topical administration may be formulated as liquid or semi-solid formulations of the present invention.

41, for example, in the form of dilute ointments, lotions, oil-in-water or water-in-oil emulsions, for example creams, ointments, pempes and pastes, including toothpastes; or in the form of solutions or suspensions such as drops, etc.

The pharmaceutical compositions of the present invention may also contain other therapeutically active compounds commonly used to treat the aforementioned conditions, such as Vitamin D ₂ and D ₃ , and hydroxylated derivatives thereof such as 1α-hydroxy-D ₃ , a-hydroxy vitamin D ₂ , lα-25-dihydroxyvitamin D ₃ , lα-25-dihydroxyvitamin D ₂ , calcitonin (of human, porcine or salmon origin), mitramycin, sodium fluoride, estrogens, and non-steroidal anti-inflammatory agents such as acetylsalicylic acid, indomethacin, naprozine and thymegadine.

The dosage of the active compounds in the compositions of the invention may vary. However, it is necessary that the amount of the active ingredient be sufficient for a suitable dosage form. The dosage selected will vary with the desired therapeutic effect, the route of administration and the duration of treatment. Dosage levels of bisphosphonate are generally from 0.001 to 10 mg / kg body weight, preferably from about 0.01 to 1.0 mg / kg. The growth hormone secretagogue is administered at a dose of 0.0001 to 25 mg / kg body weight per day to the patient for effective prevention or treatment of osteoporosis.

The combination of the invention may also be administered intermittently. For the treatment and prevention of bone resorption disorders, a typical primary oral dose of bisphosphonate is between 0.001 mg and 10 mg / kg body weight and the growth hormone secretagogue is 0.0001 to 25 mg / kg body weight. · ··· • ··· · · · · · · · · · · · · · · · · · · · · · · · ·

42 kg or a maintenance dose of one or both components, which is about half of the primary dose, can be given weekly, semi-weekly, bimonthly, monthly, bimonthly, quarterly, semiannually, annually or biennially.

The following examples illustrate the preparation of preferred compounds used in the combination product of the present invention. For the preparation of growth hormone secretagogues, see U.S. Patent Nos. 3,239,345, 4,040,979, 4,411,890, 5206235, 5,282,441, 5,310,737, 5,317,017 and 5,317,017; EP 0144230, EP 0513974, WO 94/07486, WO 94/08583, WO 94/13696 and Science 260, 1640-1643 (1993).

The invention is illustrated by the following non-limiting examples.

Example 1 A mixture of 4 molar 4-aminobutyric acid, 1.5 molar phosphoric acid and 500 ml of anhydrous chlorobenzene was heated to 100 ° C. At this temperature, 1.5 moles of phosphorus trichloride are added with vigorous stirring. The mixture is stirred at 100 ° C for 3 hours until the dense phase is completely formed and then allowed to cool. The solid was filtered, washed with a small amount of chlorobenzene and dissolved in water. The solution was refluxed for 1 hour, cooled and clarified with charcoal. The material is filtered and the product precipitated with excess hot methanol. The crude product is refluxed in 20% hydrochloric acid for 8 hours. The hydrochloric acid is removed by distillation and the residue is filtered out of water.

- We serve 43. 4-Amino-1-hydroxybutane-1,1-bisphosphonic acid is obtained in the form of a white crystalline powder having the structure (XIV) and having the following physical constants:

Analysis % C H% N% P% Found 17.88 5.62 4.93 23.94 Expected product 19.28 5.26 5.64 24.86 Calculated for product monohydrate 17.98 5.66 5.24 23.19

Example 2

3-Amino-3-methyl-N- [2,3,4,5-tetrahydro-2-oxo-1- {2 '- (1H-tetrazol-5-yl) [1,1'-biphenyl] - 4-yl] -methyl} -1H-benzazepin-3 (R) -yl] -butanamide

Step A: 3-Amino-2,3,4,5-tetrahydro-1H-1-benzazepin-2-one

9.22 g (45.6 mmol) of 3-azido-2,3,4,5-tetrahydro-1H-benzazepin-2-one prepared according to the method of Watthey et al., J. Med. Med. Chem. 28, 1511-1516 (1985)] - in 30 mL of methanol was hydrogenated for 4.5 hours, 1.0 g of 5% platinum on carbon catalyst containing metals in the presence of 2.76 ⁵ Pa T0. Celite was added and the mixture was filtered through a pad of celite. The filtrate was concentrated and allowed to stand at room temperature for 16 hours, the precipitated crystals were collected by filtration and dried in vacuo. Yield: 4.18 g (23.7 mmol, 52%). The mother liquors were diluted to 100 ml with methanol, treated with 2 g of charcoal, filtered through celite and the filtrate concentrated in vacuo to about 15 ml. The second crop was 2.02 g (11.5 mmol, 25%) of the title product. ·

- 44 we get two. The mother liquors were recycled to give a third crop of 0.88 g (5.0 mmol, 11%). Total yield 7.08 g (40.2 mmol, 88%).

1 H-NMR (200 MHz, CDCl ₃ ): 1.6 (bs, 2H), 1.80 (m, 1H),

2.55 (in, 2H), 2.88 (in, 1H), 3.42 (dd; 7Hz, 11Hz; 1H), 6.98 (d, 8Hz, 1H), 7.2 (m, 3H), 8.3 (bs, 1H).

FAB-MS: calculated for C ₁₀ H ₁₂ N ₂ O: 176; Found: 177 (M + H, 100%).

Step B: 3 (R) -Amino-2,3,4,5-tetrahydro-1H-1-benzazepin-2-one

3-amino-2,3,4,5-tetrahydro-1H-1-benzazepin-2-one (2.37 g, 13.5 mmol) and 2.02 g (13.5 mmol) of L tartaric acid is suspended in 40 ml of ethanol. The mixture is gently heated and complete dissolution with 5 ml of distilled water is added dropwise. The solution was cooled to room temperature and allowed to stand overnight. The precipitate was removed by filtration, washed with ethanol / diethyl ether (1: 1) and dried in vacuo. 1.75 g of crude L-tartrate salt are obtained. The mother liquors were evaporated to dryness in vacuo, redissolved in water (40 mL) and adjusted to pH 10-11 with solid potassium carbonate. The mixture was extracted with chloroform (6 x 20 mL), and the combined extracts were washed once with water and once with brine, dried over anhydrous potassium carbonate, filtered, and the solvents removed in vacuo. 1.29 g (7.33 mmol) of partially enriched 3 (R) -amine are obtained.

The original 1.75 g of L-tartrate salt was recrystallized twice from aqueous ethanol to give 1.03 g (3.17 mmol, 24%) of the purified L-tartrate salt, [α] _D = -212 ° (c = 1). , H ₂ O). Purified L-tartrate salt ···· ····

- Dissolve in water (20 mL) and adjust the pH to 10 - 1 with solid potassium carbonate. The mixture was extracted with chloroform (5 x 10 mL), and the combined extracts were washed with water and brine, dried over anhydrous potassium carbonate, filtered, and the solvents removed in vacuo. 522 mg (2.96 mmol, 22% overall yield) of 3 (S) -amine are obtained, [α] _D = -446 ° (c = 1, CH ₃ OH).

The remaining partially enriched 3 (R) -amine (1.29 g, 7.33 mmol) was treated with D-tartaric acid (1.10 g, 7.33 mmol) as described above and the resulting salt was recrystallized twice from aqueous ethanol.

1.20 g of purified D-tartrate salt are obtained, [α] _D = -214 ° (c = 1, H ₂ O).

The purified D-tartrate salt was dissolved in 20 mL of water and the free base was isolated as described above. 629 mg (3.57 mmol, 26% overall) of 3 (R) -amine are obtained, [α] _D = + 455 ° (c = 1, CH ₃ OH).

Step C: 2,2-Dimethylbutanedioic acid 4-methyl ester 2,2 g of dimethyl succinic acid (137 mmol) was dissolved in 200 mL of absolute methanol at 0 ° C and 2 mL of concentrated sulfuric acid was added dropwise. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 16 hours.

The mixture was concentrated to 50 ml in vacuo and 200 ml of saturated aqueous sodium bicarbonate solution was added slowly. The mixture was washed three times with hexane, the aqueous phase removed and cooled in an ice bath. The mixture was acidified to pH 2 by the slow addition of 6N hydrochloric acid and extracted eight times with diethyl ether. The combined extracts were washed with brine, dried over anhydrous magnesium sulfate, filtered, and the solvents removed in vacuo. The residue was dried at room temperature in vacuo. 14.7 g (91.8 mmol, 67%) · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · an an «best terms ..

46 viscous oils are obtained which slowly solidify on standing. 1 H NMR analysis showed the product to be a mixture of the title compound and 15% isomer of 2,2-dimethylbutanedioic acid 1-methyl ester.

The title compound NMR (200 MHz, CDC1 _3):

1.29 (s, 6H), 2.60 (s, 2H), 3.66 (s, 3H).

Nuclear Magnetic Resonance Spectrum (200 MHz, CDCl ₃ ): 1.28 (s, 6H),

2.63 (s, 2H), 3.68 (s, 3H).

Step D: Methyl 3- (Benzyloxycarbonylamino) -3-methylbutanoic acid

Step (C) for 2,2-dimethylbutanedioic acid 4-methyl ester (14.7 g, 91.8 mmol) containing 15% isomer 1-methyl ester compound in 150 mL benzene (13 mL, 9.4 g, 93 mL) triethylamine (1.01 eq., 1.00 eq.) followed by diphenylphosphoryl azide (21.8 mL, 27.8 g, 101 mmol, 1.1 eq.). After refluxing under nitrogen for 45 minutes, benzyl alcohol (19 mL, 19.9 g, 184 mmol, 2 equivalents) was added and the mixture was refluxed for 16 hours.

The mixture was cooled, filtered and the filtrate concentrated in vacuo to a minimum volume. The residue was redissolved in ethyl acetate (750 mL) and washed once with water, twice with saturated aqueous sodium bicarbonate, and once with brine. The organic phase was removed, dried over anhydrous magnesium sulfate, filtered, and the filtrate concentrated in vacuo to a minimum volume. The crude product was purified by medium pressure liquid chromatography on silica gel eluting with 4: 1 hexane / ethyl acetate. 18.27 g (68.9 mmol, 75%) of the title compound are obtained in the form of a pale yellow liquid, m.p. with a small amount of pure 3- (benzyloxycarbonylamino) -2,2-dimethylpropanoic acid methyl ester.

The title compound ^l H-NMR (200 MHz, CDC1 _3):

1.40 (s, 6H), 2.69 (s, 2H), 3.63 (s, 3H), 5.05 (s, 2H), 5.22 (broad s, 1H), 7.32 ( s, 5H).

1 H-NMR (200 MHz, CDCl ₃ ) of methyl 3- (benzyloxycarbonylamino) -2,2-dimethylpropanoic acid: 1.19 (s, 6H),

3.30 (d, 7Hz, 2H; resonance is a singlet in CD ₃ OD), 3.67 (s, 3H), 5.09 (s, 2H), 5.22 (br s, 1H; resonance not observed in CD ₃ OD), 7.3 (broad s, 5H).

Step E: 3- (Benzyloxycarbonylamino) -3-methylbutanoic acid

A solution of 18.27 g (68.9 mmol) of methyl 3-benzyloxycarbonylamino-3-methylbutanoate (Step D) in 20 mL of methanol was treated dropwise at room temperature with 51 mL (102 mmol, 1.5 eq.) ) With 2N sodium hydroxide solution. The mixture was stirred at room temperature for 16 hours, then transferred to a separatory funnel and washed three times with hexane. The aqueous phase was removed, cooled to 0 ° C and slowly acidified to pH 2 by dropwise addition of 6 N hydrochloric acid (indicator paper). This mixture was extracted six times with diethyl ether, and the combined extracts were washed with 1 N hydrochloric acid and brine, dried over anhydrous magnesium sulfate, filtered, and the solvent removed in vacuo. 17.26 g (68.7 mmol, 99%) of the title compound are obtained.

1 H-NMR (200 MHz, CDCl ₃ ): 1.42 (s, 6H), 2.77 (s, 2H), 5.06 (s, 2H), 5.2 (bs, 1H), δ , 3 (s, 5H).

• · · · * "······" • · · · · · · · · · · · • «• · · · ·« · · · · ···· ·· ·· * _Β

- Step F: 3- (Benzyloxycarbonylamino) -3-methyl-N- [2,3,4,5-tetrahydro-2-oxo-1H-benzazazepine-3 (R) -il 1-buta namid

A solution of 3 (R) -amino-2,3,4,5-tetrahydro-1H- [1] benzazepin-2-one (252 mg, 1.43 mmol) in 4 mL of methylene chloride at room temperature 3- (Benzyloxycarbonylamino) -3-methyl-butanoic acid (Step E) (mg) (1.60 mmol, 1.1 equiv) followed by benzotriazole (760 mg, 1.7 mmol, 1.2 equiv) -1-Oxy-tris (dimethylamino) -phosphonium hexafluorophosphate and diisopropylethylamine (0.50 mL, 380 mg, 2.9 mmol, 2 equiv.) Were added. After 3 hours at room temperature, the mixture was diluted with ethyl acetate (30 mL) and washed with 5% aqueous citric acid, saturated aqueous sodium bicarbonate (twice), and brine. The organic layer was removed, dried over anhydrous magnesium sulfate, filtered and the solvents removed in vacuo. The residue was purified by medium pressure liquid chromatography on silica gel eluting with ethyl acetate. 586 mg (1.43 mmol, 100%) of the title product are obtained. 1 H-NMR (200 MHz, CDCl ₃ ): 1.38 (s, 3H), 1.39 (s, 3H), 1.82 (m,

1H), 2.52 (s, 2H), 2.5-3.0 (m, 3H), 4.51 (m, 1H), 5.07 (broad s, 2H), 5.57 (broad s) , 1H), 6.68 (d, 7Hz, 1H), 6.97 (d, 8Hz, 1H), 7.1-7.4 (m, 8H), 7.61 (broad s, 1H).

FAB-MS: calculated for C ₂₃ H ₂₇ N ₃ O ₄ : 409; Found: 410 (M + H, 100%);

[α] _D = + 137 ° (c = 1, CHCl ₃ ).

Step G: To 5 mL of 5-phenyltetrazole in ml of N, N-dimethylformamide was added small portions of 3.3 g (24.3 mmol, 0.5 equiv.) Of zinc chloride while maintaining the temperature. · · · · · · · · · · · · · · · · · · · · · · ···

- 49 feces maintained below 60 ° C. The zinc chloride suspension was cooled to room temperature and treated with 5.0 g (48.5 mmol, 1.0 equivalent) of benzonitrile followed by 3.2 g (48.5 mmol, 1.0 equivalent) of sodium azide. The heterogeneous mixture was heated at 115 ° C with stirring for 18 hours. The mixture was cooled to room temperature, water (30 ml) was added and the mixture was acidified by addition of concentrated hydrochloric acid (5.1 ml). The mixture was cooled to 0 ° C and allowed to stand for 1 hour, filtered and the precipitate was washed with cold 0.1 N hydrochloric acid (15 mL) and dried in vacuo at 60 ° C. 6.38 g (43.7 mmol, 90%) of the title product are obtained.

Step H: 5-Phenyl-2-trityltetrazole

To a suspension of 5-phenyltetrazole (5.0 g, 34.2 mmol) in acetone (55 mL) was added triethylamine (5.0 mL, 3.6 g, 35.6 mmol, 1.0 equivalent). After 15 minutes, a solution of triphenylmethyl chloride (10.0 g, 35.9 mmol, 1.05 equiv.) In tetrahydrofuran (20 mL) was added and the mixture was stirred at room temperature for 1 hour. Water (75 mL) was added slowly and stirred at room temperature for 1 hour. The product was collected by filtration, washed with water (75 mL) and dried in vacuo at 60 ° C. 13.3 g (34.2 mmol, 100%) of the title product are obtained.

Step I: N- (Triphenylmethyl) -5- [2- (4'-methylbiphen-4-yl)] - Tetrazole

A solution of zinc chloride (6.3 g, 46.2 mmol, 0.6 equiv.) In tetrahydrofuran (35 mL) was dried over molecular sieve.

5-Phenyl-2-trityltetrazole (30.0 g, 77.3 mmol, 1.0 eq.) Was dissolved in anhydrous tetrahydrofuran (300 mL) and the solution was gently stirred while degassing three times with alternating vacuum and nitrogen purge. The stirred solution was cooled to -15 ° C and ······ ··· «··

50 ml of a 1.6 mol / l solution of n-butyllithium in hexane (80.0 mmol, 1.05 equivalents) are slowly added while maintaining the temperature below -5 ° C. After 1.5 hours at -5 to -15 ° C, the mixture was treated with the dried zinc chloride solution and allowed to warm to room temperature.

In another flask, 20.17 g (92.5 mmol, 1.2 equivalents)

4-iodo-toluene and bis (triphenylphosphine) nickel (II) dichloride (1.5 g, 2.3 mmol, 0.03 equiv.) Were dissolved in tetrahydrofuran (60 mL), degassed and left under nitrogen. The mixture was cooled to 5 ° C and treated with 1.5 ml of 3.0M magnesium chloride in THF (4.5 mmol, 0.06 equiv) while keeping the temperature below 10 ° C. we. The solution was warmed to room temperature and added to the arylzinc solution under a stream of nitrogen. After vigorously stirring at room temperature for 8 hours, the reaction was quenched by slow addition of a solution of glacial acetic acid (10 mL, 1.6 mmol, 0.02 equivalent) in tetrahydrofuran (60 mL) at a rate below 40 ° C. . The mixture was stirred for 30 minutes and 150 ml of 80% saturated aqueous sodium chloride solution was added; the reaction mixture is extracted for 30 minutes and the phases are separated. The organic phase is removed and washed with 150 ml of 80% aqueous sodium chloride solution buffered with ammonium hydroxide to pH> 10. The organic phase was removed and concentrated in vacuo to about 50 mL and then acetonitrile (250 mL) was added. The mixture was again concentrated in vacuo to 50 ml and the volume was adjusted to 150 ml with acetonitrile. The resulting suspension was cooled to 5 ° C for 1 hour, then filtered and washed with cold acetonitrile (50 mL) and distilled water (150 mL). The precipitate was air dried and the resulting free flowing solid was then dried at 50 ° C in vacuo for 12 hours. 30.0 g (62.8 mmol, 81%) of product are obtained.

1 H-NMR (200 MHz, CDCl ₃ ): 2.28 (s, 3H), 6.9-7.05 (m, 10H),

7.2-7.5 (m, 12H); 7.9 (m, 1H).

Step J: N- (Triphenylmethyl) -5- {2- {4 '- (bromomethyl) biphen-4-yl]} tetrazole

A solution of N-triphenylmethyl-5- [2- (4'-methylbiphen-4-yl)] - tetrazole (3.15 g, 6.6 mmol) in methylene chloride (25 mL) was added. Treatment with 29 g (7.25 mmol, 1.1 equivalents) of N-bromosuccinimide, 80 mg (0.5 mmol, 0.07 equivalent) of AIBN, 200 mg of sodium acetate and 200 mg of acetic acid. The mixture was refluxed for 2 to 16 hours, then cooled and washed with saturated aqueous sodium bicarbonate. The organic layer was removed, dried over anhydrous sodium sulfate, filtered, and concentrated to a minimum volume by atmospheric distillation. Methyl tert-butyl ether is added and distillation is continued until almost all methylene chloride is removed and the total volume is reduced to about 12 ml, at which time 12 ml of hexane are added. After 2 hours at room temperature, the product was isolated by filtration, washed with hexanes and dried in vacuo at 50 ° C. Yield: 2.81 g (5.04 mmol, 76%). 1 H-NMR (200 MHz, CDCl ₃ ): 4.38 (s, 2H), 6.9-8.0 (m, 23H).

• · · · ····

- 52 NMR data shows about 1% starting material and 7% dibromo derivative.

Step K: 3- (Benzyloxycarbonylamino) -3-methyl-N- {2,3,4,5-tetrahydro-2-oxo-1 - [{2 '- [N- (triphenyl) methyl) -tetrazul-5-yl] - [1,1'-biphenyl] -4-yl} methyl] -1H-1-benzazepin-3 (R) -yl} -butanamide

To a solution of the intermediate obtained in Step F above (437 mg, 1.07 mmol) in dry dimethylformamide (mL) at room temperature under nitrogen atmosphere was added 55 mg of 60% sodium hydride in oil (33 mg NaH, 1.38 mmol, 1.3 equivalents). We are added. After 15 minutes, 715 mg (1.28 mmol, 1.2 equivalents) of N-triphenylmethyl-5- [2- (4'-bromomethyl-biphen-4-yl)] - tetrazole (Step J) is added. A solution of 1.5 ml of anhydrous dimethylformamide was added and the mixture was stirred for 90 minutes.

The reaction mixture was added to ethyl acetate (100 mL) and washed with water (twice) and brine. The organic layer was removed, dried over anhydrous magnesium sulfate, filtered and the solvents removed in vacuo. Purification by silica gel medium pressure chromatography, eluting with 1: 1 ethyl acetate: hexane gave 902 mg (1.02 mmol, 95%) of product.

1 H-NMR (200 MHz, CDCl ₃ ): 1.38 (s, 3H), 1.39 (s, 3H), 1.68 (m,

1H), 2.2-2.5 (m, 5H), 4.44 (m, 1H), 4.67 (d, 14Hz, 1H),

5.06 (s, 2H), 5.12 (d, 14Hz, 1H), 5.63 (broad 1, 1H), 6.65 (d, 8Hz, 1H), 6.9-7.5 (m , 31H), 7.85 (m, 1H).

· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

- 53 Step L: 3-Amino-3-methyl-N- [2,3,4,5-tetrahydro-2-oxo-1- {1,2 '- (1H-tetrazol-5-yl)]. l, l'-biphenyl | -4-yl] methylethoxyimino} -lH-l-benzazepin-3 (R) -yl] butanamide, trifluoroacetate

A solution of the intermediate obtained in Step H above (902 mg, 1.02 mmol) in methanol (mL) was hydrogenated at room temperature and pressure (9.8 bar) over 20 mg of 20% Pd (OH) ₂ / C catalyst for 14 hours. The mixture was filtered through celite and concentrated in vacuo. The residue was purified by reverse phase HPLC on C-18-η, eluting with methanol / 0.1% aqueous trifluoroacetic acid (linear gradient from 60% methanol to 80% over 10 min). 568 mg (0.91 mmol, 89%) of the title compound are obtained.

1 H-NMR (200 MHz, CD ₃ OD): 1.33 (s, 3H), 1.37 (s, 3H), 2.0-2.6 (m, 6H), 4.35 (dd; 7, 11Hz; 1H), 4.86 (d, 15Hz, 1H), 5.20 (d, 15Hz, 1H), 7.00 (d, 8Hz, 2H), 7.15-7.35 (m , 6H), 7.45-7.70 (m, 4H).

FAB-MS: calculated for C ₂₉ H ₃₁ N ₇ O ₂ : 509; found: 510 (M + H, 100%).

Example 3

3 - [(2 (R) -Hydroxypropyl) amino] -3-methyl-N- [2,3,4,5-tetrahydro-2-oxo-l - {[2 '- (lH-tetrazol 5-yl) [1,1'-Biphenyl] -4-yl] methyl} -1H-benzazepin-3 (R) -yl] butanamide

Step A: 3 - {[2 (R) - (Benzyloxy) -propyl] -amino} -3-methyl-N- [2,3,4,5-tetrahydro-2-oxo-1 - {[ 2 '- (tetrazol-5-yl) [l, l'-biphenyl] -4-yl] methyl} -lH-l-benzazepin-3 (R) -yl] butanamide, trifluoroacetate

The title compound is 3-amino-3-methyl-N- [2,3,4,5-tetrahydro-2-oxo-1 - {[2 '- (1H-tetrazol-5-yl) [1,1] '-biphenyl] -4-yl] -methyl} • · ··· · · · · · · · · · · ·

- 54 -1 Η -1 - Nazazepine-3 (R) -1-butanamido-d-trifluoroacetate (Example 1) and (R) -2-benzyloxy- propanal, prepared from ethyl D-lactate according to the procedure of Hanessian and Kloss (Tetrahedron Lett., 26, 1261-1264 (1985)), by the method described in Example 86 (A) of US 5206235.

1 H-NMR (200 MHz, CD ₃ OD): 1.25 (d, 6Hz, 3H), 1.35 (s, 6H),

2.11 (m, 1H), 2.32 (m, 1H), 2.5-2.7 (m, 4H), 2.95 (m, 1H),

3.17 (m, 1H), 3.80 (m, 1H), 4.40 (m, 1H), 4.44 (d, 11Hz,

1H), 4.64 (d, 11Hz, 1H), 4.90 (d, 15Hz, 1H), 5.02 (d, 15Hz,

1 H), 6.99 (d, 8 Hz, 2H), 7.1-7.7 (m, 15H).

Calcd for C ₃₉ H ₄ 3 N ₇ O _3: Calcd: FAB-MS 657; found: 658 (M + H, 100%).

Step B and 3 - [(2 (R) -Hydroxypropyl) amino] -3-methyl-N- (2,3,4,5-tetrahydro-2-oxo-1 - {[2'- (1H-Tetrazol-5-yl) [1,1'-biphenyl] -4-yl] methyl} -1H-1-benzazepin-3 (R) -yl] -b followed by trifluoroacetate

The title compound is prepared from the intermediate obtained in Step A according to the procedure described in Example 86, step B of US 5206235.

1 H-NMR (400 MHz, CD 3 OD): 1.22 (d, 6Hz, 3H), 1.37 (s, 3H),

1.39 (s, 3H), 2.10 (m, 1H), 2.31 (m, 1H), 2.45-2.70 (m, 4H),

2.81 (dd; 10, 12Hz; 1H), 3.08 (dd; 4, 12Hz; 1H), 3.92 (m,

1H), 4.36 (dd; 7, 11Hz; 1H), 4.93 (d, 15Hz, 1H), 5.17 (d,

15Hz, 1H), 7.04 (d, 8Hz, 2H), 7.19 (d, 8Hz, 2H), 7.20-7.35 (m, 4H), 7.54 (m, 2H), 7 , 65 (in, 2H).

FAB-MS: calculated for C ₃₂ H ₃₇ N ₇ O ₃ : 567; found: 568 (M + H, 45%).

• · ···· ···· · ·

- 55 Example 4 (Method 1)

N | l (R) - [(l, 2-dihydro-l-methanesulfonyl-spiro | 3H-indole-3,4'-piperidin-l-yl) -karboniIl-2- (phenyl-methyl) -etill- 2-amino-2-methyl-propánaniid

Step A: 1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidine] hydrochloride

1.20 g (5.8 mmol) of 1'-methyl-1,2-dihydro-spiro [3H-indole-3,4'-piperidine] (prepared according to the procedure of H. Ong et al., J. Med. Chem. 23, 981-986 (1983)] in 20 ml of anhydrous dichloromethane at 0 ° C were treated with 0.90 ml (6.4 mmol) of triethylamine and 0.49 ml (6.35 mmol) of methanesulfonyl chloride. and the reaction mixture was stirred for 30 minutes. The reaction mixture was poured into saturated aqueous sodium bicarbonate solution (15 mL) and extracted with dichloromethane (2 x 10 mL). The combined organic extracts were washed with brine (20 mL), dried over anhydrous potassium carbonate, filtered and the solvent removed in vacuo. 1.44 g of the methanesulfonamide derivative are obtained in the form of a pale yellow oil which is used without purification.

To a solution of the above crude product in 20 ml of anhydrous 1,2-dichloroethane was added 1.0 ml (9.30 mmol) of (1-chloroethyl) chloroformate at 0 ° C and the mixture was stirred at room temperature for 30 minutes. stir and finally reflux for 1 hour. The reaction mixture was concentrated to about one-third volume, diluted with anhydrous methanol (20 mL) and refluxed for 1.5 hours. The reaction mixture was cooled to room temperature and concentrated to about half its volume. The precipitate was filtered off and washed with a small amount of methylene chloride. 1.0 g of the title piperidine hydrochloride salt are obtained in the form of a white solid. The filtrate was concentrated and a little methanol was added followed by diethyl ether. The precipitated material is filtered off again, washed with cold methanol and dried. An additional 0.49 g of the desired product is obtained. Total yield 1.49 g (70%).

1 H-NMR (CDCl ₃ 200 MHz): 7.43-7.20 (m, 3H), 7.10 (dd, 1H),

3.98 (bs, 2H), 3.55-3.40 (bd, 2H), 3.35-3.10 (m, 2H),

2.99 (s, 3H), 2.15 (t, 2H), 2.00 (t, 2H).

Step B: N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) -carbonyl] -2-phenyl methyloxy) ethyl} -2 - [(l, l-dimethylethoxy) carbonylamino] -2-methylpropanamide

0.35 g (1.15 mmol) of (2R) -2 - [(1,1-dimethylethoxy) carbonylamino] -3- [2- (phenylmethyloxy) ethyl] -1- propanoic acid and 13 mL of dichloromethane, 0.325 g (1.07 mmol) of 1,2-dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidine] hydrochloride, 0.18 mL (1.63 N-methylmorpholine (0.1 mmol) and 1-hydroxybenzotriazole (HOBT) (0.159 g, 1.18 mmol) were added and the mixture was stirred for 15 minutes. EDC (0.31 g, 1.62 mol) was added and stirring was continued for 1 hour. Another 60 µl of N-methylmorpholine was added and stirred for 45 minutes. The reaction mixture was poured into water (5 mL) and the organic layer was separated. The organic layer was washed with 5 mL of 0.5 N aqueous hydrochloric acid and 5 mL of saturated aqueous sodium bicarbonate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. 0.627 g of product is obtained in the form of a yellow foam which is used without further purification.

• ·

To a mixture of the above product (0.627 g, 1.07 mmol) and dichloromethane (5 mL) was added trifluoroacetic acid (1.0 mL) and stirred at room temperature for 75 minutes. After adding another 1.00 ml of trifluoroacetic acid, the mixture was stirred for 10 minutes. The reaction mixture was concentrated, diluted with dichloromethane (5.0 mL) and carefully basified by pouring into 10 mL of 10% aqueous sodium carbonate. The organic layer was separated and the aqueous layer was further extracted with dichloromethane (2 x 15 mL). The combined organic layers were washed with water (5 mL), dried over anhydrous potassium carbonate, filtered and concentrated. 0.486 g of amine is obtained in the form of a pale yellow foam which is used without further purification.

To a mixture of the above amine (0.486 g, 1.01 mmol) and dichloromethane (10 mL) was added 2 - [(1,1-dimethylethoxy) carbonylamino] -2-methylpropanoic acid (0.26 g, 1.28 mmol). 1-hydroxybenzotriazole (HOBT) (0.173 g, 1.28 mmol) and EDC (0.245 g, 1.28 mol) were added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into 5.0 mL of water and the organic layer was separated. The aqueous phase was back-extracted with dichloromethane (5 mL). The combined organic layers were washed with 5.0 mL of 0.5 N aqueous hydrochloric acid, 5 mL of saturated aqueous sodium bicarbonate, dried over anhydrous magnesium sulfate, and then concentrated. 0.751 g of crude product is obtained in the form of a yellow foam. A solution of the above crude product in dichloromethane is chromatographed on silica gel (25 g), eluting first with 70/25/5 hexanes / acetone / dichloromethane and then 65/30/5 hexanes / acetone / dichloromethane. 0.63 g of the title compound is obtained in the form of a white solid.

···· ····

- 58 H-NMR (CDCl _3, 400 MHz) δ: 7.40-7.10 (m, 6H), 7.06 (d, 1 / 3H),

7.02 (t, 1 / 3H), 6.90 (t, 1 / 3H), 6.55 (d, 1 / 3H), 5.15 (m, 1H),

4.95 (br s, 1H), 4.63 (br d, 1 / 3H), 4.57-4.40 (m,

2/3 H), 4.10 (br d, 1 / 3H), 4.00 (br d, 1 / 3H), 3.82 (t, 1H), 3.78-3.62 (m, 2H) ), 3.60-3.50 (m, 1H), 3.04 (q, 1H),

2.87 (s, 1H), 2.86 (s, 2H), 2.80-2.60 (m, 1H), 1.90 (broad s,

1H), 2.85-2.75 (m, 1H), 1.82-1.60 (m, 3H), 1.55-1.45 (m,

1H), 1.45 (s, 4H), 1.42 (s, 2H), 1.39 (s, 9H).

Step C: N- [1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) -carbonyl-2- (phenyl) -methyloxy) ethyl] -2-amino-2-methylpropanamide hydrochloride

To a mixture of the intermediate prepared in Step B above (0.637 g, 0.101 mmol) and dichloromethane (5 mL) was added trifluoroacetic acid (2.5 mL) and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated and the resulting oil was taken up in 10 mL of ethyl acetate and washed with 8 mL of 10% aqueous sodium carbonate. The aqueous phase was further extracted with ethyl acetate (5 mL). The combined organic extracts were washed with water (10 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated. 0.512 g of the title compound is obtained as the free base in the form of a white foam.

To a mixture of 0.512 g of the free base and 5 ml of ethyl acetate was added ethyl acetate saturated with 0.2 ml of hydrochloric acid and the mixture was stirred for 1.5 hours. The resulting white precipitate was filtered under nitrogen, washed with diethyl ether and dried. 0.50 g of the title compound is obtained in the form of a white solid.

1 H-NMR (400 MHz, CD ₃ OD) δ: 7.40-7.28 (in, 4H), 7.25-7.17 (m, 2H), 7.08 (t, 1 / 3H) , 7.00 (t, 1 / 3H), 6.80 (d, 1 / 3H), 5.16

- 59 ······································································································································································································································· · ddd, 1H), 4.60-4.42 (in, 3H), 4.05 (t, 1H), 3.90 (broad s,

2H), 3.83-3.70 (m, 2H), 3.30-3.15 (m, 1H0, 2.97 (s, 1H),

2.95 (s, 2H), 2.90-2.78 (in, 1H), 1.96 (t, 1 / 3H), 1.85-1.65 (in, 4H), 1.63 ( s, 2H), 1.60 (s, 4H).

The compound is obtained as a 3/2 mixture of rotamers.

Example 5 (Method 2)

N- {l (R) - [(l, 2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4 ^'- P> P ^er pyrrolidin] -l'-yl) carbonyl] -2- ( phenyl-methyl) ethyl} -2-amino-2-methylpropanamide

Step A: (2R) - [[[2- (1,1-Dimethylethoxy) carbonyl] amino] -2,2-dimethyl-1-oxoethyl] amino-2- (phenylmethoxy) ethyl] -l-propanoic acid allyl ester

The title compound is prepared from (2R) -2 - [(1,1-dimethylethoxy) carbonylamino] -3-phenylmethyloxyethylpropanoic acid and allyl alcohol, the coupling reaction being carried out in dichloromethane. -methane in the presence of EDC and DMAP.

1 H-NMR (400 MHz, CDCl ₃ ) δ: 7.25 (s, 5H), 5.8 (m, 1H), 5.2 (dd, 2H), 5.0 (bs, 1H), 4.7 (m, 1H), 4.6 (in, 2H), 4.4 (dd, 2H), 3.9 (dd, 1H), 3.6 (dd, 1H), 1.45 (d) , 6H), 1.39 (s, 9H).

Step B: (2R) - [{[2- (1,1-Dinethylethoxy) carbonyl] amino} -2,2-dimethyl-1-oxoethyl] amino-2- (phenylmethyl) oxy) ethyl) -1-propanoic acid

6.7 g (15.9 mmol) of the crude product from Step A above, 1.8 g (0.1 equivalent) of tetrakis (triphenylphosphine) palladium and 1.25 g (0.3 equivalent) of triphenyl- To the solution of phosphine was added 35 ml of a 0.5 M solution of potassium 2-ethylhexanoate in ethyl acetate with stirring. The reaction mixture room • · ·············································································································································.

After stirring at 60 ° C for 1 hour under nitrogen, it was diluted with diethyl ether (100 mL) and poured into ice water. The organic layer was separated and the aqueous layer was acidified with 20% citric acid and extracted with ethyl acetate. The ethyl acetate extracts were washed with brine, dried over anhydrous magnesium sulfate, filtered and evaporated. The title compound is obtained as a solid.

1 H-NMR (400 MHz, CD ₃ OD) δ: 7.3 (s, 5H), 4.7 (m, 1H), 4.5 (s,

2H), 4.0 (m, 1H), 3.6 (m, 1H), 1.4 (d, 6H), 1.3 (s, 9H).

Step C: N- [1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin-1'-yl) carbonyl] -2- (phenyl) methyloxy) ethyl] -2- (l, l-dimethylethoxy) carbonyl-amino-2-methylpropanamide

1.0 g (3.44 mmol) of 1-methanesulfonyl-spiro [indoline-3,4'-piperidine] hydrochloride, 1.44 g (3.78 mmol) of (2R) - {[2- (1,1-) dimethylethoxy-carbonyl] -amino} -2,2-dimethyl-1-oxoethyl] -amino-2- (phenylmethyloxy) ethyl) -1-propanoic acid, 0.58 ml (5, To a solution of N-methylmorpholine (20 mmol) and 1-hydroxybenzothiazole (HOBT) (0.58 g, 3.78 mmol) in dichloromethane (50 mL) was added EDC (1.03 g, 5.20 mmol), and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with another 50 ml of dichloromethane, washed with 50 ml of aqueous sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered and concentrated. The crude oily residue was purified by flash chromatography on silica gel (50 g) to give the title compound (2.148 g, 90%) as a colorless foam.

1 H NMR (CDCl _3, 400 MHz,) δ: 7.40-7.10 (m, 6H), 7.06 (d,

1 / 3H), 7.02 (t, 1 / 3H), 6.90 (t, 1 / 3H), 6.55 (d, 1 / 3H), 5.15

- 61 • · · · ···· (in, 1 Η), 4.95 (wide s, 1H), 4.63 (wide d, 1 / 3H), 4.57-4.40 (m, 2 2/3 H), 4.10 (broad d, 1/3 H), 4.00 (broad d,

1/3 H), 3.82 (t, 1H), 3.78-3.62 (m, 2H), 3.60-3.50 (m, 1H),

3.04 (q, 1H), 2.87 (s, 1H), 2.86 (s, 2H), 2.80-2.60 (m, 1H),

1.90 (br s, 1H), 2.85-2.75 (m, 1H), 1.82-1.60 (m, 3H),

1.55-1.45 (m, 1H), 1.45 (s, 4H), 1.42 (s, 2H), 1.39 (s, 9H).

The compound was obtained as a 3/2 mixture of rotamers.

D) Preparation of N- {l (R) - [(l, 2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4'-pip ^er id * n] -l'-yl) carbonyl] -2- (phenyl-methyl) ethyl} -2-amino-2-methylpropanamide hydrochloride

To a solution of the intermediate prepared in Step C above (2.148 g, 3.41 mmol) in dichloromethane (10 mL) was added trifluoroacetic acid (5 mL) and stirred for 1 hour. The reaction mixture was concentrated and made basic with 100 ml of 5% aqueous sodium carbonate solution and extracted with 3 x 50 ml of dichloromethane. The combined organic layers were washed with brine (50 mL), dried over anhydrous potassium carbonate, filtered and concentrated. The resulting colorless foam was dissolved in ethyl acetate (25 mL), cooled to 0 ° C, and treated with ethyl acetate (4 mL, 1 M in ethyl acetate). The precipitate was filtered and washed first with ethyl acetate and then with ethyl acetate / diethyl ether (1: 1) and dried. Obtained as a colorless solid (1.79 g, 93%).

1 H-NMR (400 MHz, CD ₃ OD) δ: 7.40-7.28 (m, 4H), 7.25-7.17 (m, 2H), 7.08 (t, 1 / 3H) , 7.00 (t, 1 / 3H), 6.80 (d, 1 / 3H), 5.16 (ddd, 1H), 4.60-4.42 (m, 3H), 4.05 (t , 1H), 3.90 (s broad,

2H), 3.83-3.70 (m, 2H), 3.30-3.15 (m, 1H0, 2.97 (s, 1H),?

- 62 2.95 (s, 2H), 2.90-2.78 (m, 1H), 1.96 (t, 1 / 3H), 1.85-1.65 (m, 4H), 1, 63 (s, 2H), 1.60 (s, 4H).

The compound was obtained as a 3/2 mixture of rotamers.

Example 6

N- {l (R) - [l, 2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -l'-yl) carbonyl-3-phenyl-propyl} -2- amino-2-methylpropanamide hydrochloride

Step A: N- {1 (R) -1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl- propyl} -2 - [(1,1-dinethyl-ethoxy) -carbonyl-amino] -2-methyl-piperaldehyde

The title compound was obtained from (2R) -2 - [(1,1-dimethylethoxy) carbonylamino] -4-phenyl-1-butanoic acid and 1,2-dihydro-1-methylsulfonyl-spiro [3H- indole-3,4'-piperidine] hydrochloride a

Example 18 by the coupling procedure described in Step B). The crude product was purified on silica gel eluting with 5% acetone in dichloromethane.

¹ H-NMR (400 MHz, CDCl ₃ ) δ: 7.2 (m, 9H), 4.9 (m, 1H), 4.5 (m,

1H), 3.8 (m, 2H), 3.2 (m, 2H), 2.9 (s, 3H), 2.7 (m, 2H), 2.3 (s, 2H), 2, 0 (m, 2H), 1.7 (m, 4H), 1.5 (s, 6H), 1.4 (s, 9H).

Step B: N- {1 (R) - [1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) -carbonyl} -3-phenyl- propyl} -2-amino-2-methylpropanamide hydrochloride

The title compound was prepared from the intermediate obtained in Step A above using the deprotection procedure described in Example 18, Step C.

1 H-NMR (400 MHz, CD ₃ OD) δ: 7.3 (m, 9H), 4.5 (m, 1H), 3.9 (m, 2H), 3.5 (m, 2H), 3.2 (m, 2H), 2.9 (s, 3H), 2.7 (m, 4H),

- 63 2.0 (in, 4H), 1.6 (s, 6H).

Example 7

N- {l (R) - [(l, 2-dihydro-l-methanesulfonyl-spirol3H-indole-3,4'-piperidin] -l'-yl) carbonyl-3-phenyl-propyl} -2-Amino Combination therapy with -2-methylpropanamide and pamidronate: exploratory nine-week bone analysis in old female rats

The purpose of the study was to evaluate the N- {1 (R) - [(1,2-dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] ] -3-phenylpropyl} -2-amino-2-methylpropanamide, alone or in combination with pamidronate (3-amino-1-hydroxypropylidene-1,1-bisphosphonic acid) on bone of old female rats. The duration of the study was 9 weeks. N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} Frequency of administration of -2-amino-2-methylpropanamide: once daily, weekly. Frequency of dosing of pamidronate: once a week, on the first day of the week. N- {1 (R) - [(1,2-Dihydro-1-methanesulfonylspiro [3 H -indo-3,4,4'-piperidin] -1'-yl) carbonyl] -3-phenylpropyl} -2-Amino-2-methylpropanamide was administered by probe. As a control, distilled water was used and the vehicle was also distilled water. N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} -2 -amino-2-methylpropanamide has a dose volume of 5 ml / kg and pamidronate has a dose volume of 1 ml / kg. The study was performed on female rats of the Sprague-Dawley Crl: CD® (SD) BR strain, which were more than 18 months old at the start of the study, and

64 weighing approximately 300-400 g at the start of the study.

The food and water did not contain any impurities that were known to interfere with the study. The litter did not contain any contaminants known to interfere with the test. The rats were individually housed in stainless steel wire cages.

Dose Levels Dose Code Number of Animals

Male Female

Control 1 01

Controls

Distilled water 07

N- {1 (R) - [(1,2-dihydro-1-15)

-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} -2-amino-2-methyl-propanamide [50 mg / kg / day]

Pamidronate 25 [120 pg / kg / weekly]

N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl- propyl} -2-amino-2-methylpropanamide + Pamidronate [50 mg / kg / day +

120 pg / kg / weekly]

7

8

11

10 • · · · • ·

- 65 Hormone Analysis

First day of drug delivery: Blood samples (approximately 1.5 ml) were taken from the ocular cavity of non-fasted rats in each group to determine GH; blood sampling in the control group and N- {1 (R) - [(1,2-dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] - 15 minutes after dosing, 3-phenylpropyl} -2-amino-2-methylpropanamide was administered alone or in combination.

Medication weeks 2 and 9:

Blood samples (about 1.5 mL) were taken from the ocular cavity of non-fasted rats in all groups except control group 1 for GH determination; blood control in control group 2 and N- {1 (R) - [(1,2-dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] 3-phenylpropyl} -2-amino-2-methylpropanamide, alone or in combination, 15 minutes after dosing; blood samples (maximum volume passage) were obtained from venous cava at necropsy in non-fasted rats for determination of IGF-1 (* only for control group 1).

· «· ·« • · · · · ·

Growth hormone levels (1 day)

Control 1 10 ng / ml

Control 2 10 ng / ml

Pamidronate 5 ng / ml

N- {1 (R) - [(1,2-Dihydro-1-methane-85 ng / ml sulfonyl spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3 phenyl-propyl} -2-amino-2-methyl-propánainid

N- {1 (R) - [(1,2-Dihydro-1-methane-48 ng / ml sulfonylspiro [3H-indo-3,4,4'-piperidin] -1'-yl) carbonyl] - 3-phenylpropyl} -2-amino-2-methylpropanamide + pamidronate

bone Forecast

All rats received bone markers (oxytetracycline and calcein): oxytetracycline for 9 days and calcein for 2 days before autopsy.

Oxytetracycline was injected subcutaneously twice (approximately 5 hours between the two injections) at 25 mg / kg and calcein was injected intraperitoneally at 15 mg / kg.

- 67 • · '• ··

Effect on the surface of bone-forming cells Treatment Bone-forming cell surface% (± SEM)

Controls

N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} - 2-amino-2-methylpropanamide

pamidronate

N- {l (R) - [(l, 2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -l'-yl) carbonyl] -3-phenyl-propyl} - 2-amino-2-methylpropanamide + pamidronate

3.51 (0.63) 3.91 (0.73)

0.80 (0.26) 3.05 (0.36)

As indicated by the above results, the growth hormone secretagogue restores bone formation, which was suppressed to the control level by bisphosphonate-type pamidronate. In addition, there was no difference in the surface of bone-degrading cells (bone resorption) as a result of treatment with a growth hormone secretagogue. The results of the study reflect the unexpected advantage that can be achieved with the present invention.

Although the invention has been illustrated by specific embodiments, it will be apparent to those skilled in the art that various adaptations, changes, modifications, substitutions, deletions, or substitutions may be made to the methods and protocols without departing from the spirit of the invention. For example, ···· ···· · · ·

Effective doses other than those indicated may be used because the response of the treated mammal to treatment may vary with any of the compounds of the invention. Similarly, the specific pharmacological responses observed may vary depending upon the specific active agent being treated or the pharmaceutical carriers present, and the type of formulation used and the mode of administration, and therefore such expected changes or differences in results are still within the scope of the present invention. and in accordance with the embodiment thereof. The invention is defined by the scope of the following claims, which are to be construed in the broadest possible sense.

Claims (18)

Claims - 11) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl- propyl} -2-amino-2-methylpropanamide; 1) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (1H indol-3-yl} ethyl) -2-amino-2-methylpropanamide; • · · · · · · · · · · · · · · · · · · · · · · · · ··· - 85 2) Ν - {1 (R) - [(1,2-Dihydro-1-methanecarbonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2 - (1H-indol-3-yl) -ethyl} -2-amino-2-methyl-propanamide; C 1-3 (C 1 -C 10) alkanoyloxy, C 1-3 (C 1 -C 6) alkoxy, phenyl, phenoxy, 2-furyl, (C 1-6) alkoxycarbonyl, (1-6) C 1) alkyl substituted with -O _m -S; obsession R ₄ and R _{5 taken} together may form - (CH ₂ ) _r L _a (CH ₂ ) _s -, wherein L _a is -C (R ₂ ) ₂ -, -O-, -S (O) _m - or -N (R ₂ ) -, wherein r and s are independently 1-3 and R ₂ is as defined above; R ₆ is hydrogen or C 1-6 alkyl; A is a group of formula (a) or (b) wherein x and y are independently 0-3, R ₇ and R _7a are each independently hydrogen, C 1-6 alkyl, -OR ₂ , trifluoromethyl, phenyl, substituted C 1-6 alkyl, wherein the substituents are imidazolyl, phenyl, indolyl, p-hydroxyphenyl; -OR ₂ , 1-3 fluorine atoms, Can be -S (O) _m R _2, -C (0) 0R _2, C3-7 cycloalkyl, N _{(R2) (R2),} -C (O) N _{(R2) (R2)} are: , obsession 1-9 halogens, S (O) _m R _2a , 1-3 OR _2a · « - may contain 80 groups or C (O) OR _2a and the aryl groups may be further substituted with phenyl, phenoxy, halophenyl, C 1-3 alkyl, 1-3 halogen, 1-2-OR ₂ , methylenedioxy group, -S (O) R ₂ group _female, 1-2 _CF3 groups, -OCF _3, -NO _2, -N _{(R2) (R2),} -N (R ₂ ) C (O) R ₂ , -C (O) OR ₂ , -C (O) N (R ₂ ) (R ₂ ), -SO ₂ N (R ₂ ) (R ₂ ), - They may contain N (R ₂ ) S (O) ₂ -aryl and -N (R ₂ ) SO ₂ R ₂ ; R ₂ is hydrogen, C 1-6 alkyl, 1) nh ₂ , 1) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -2- (1H) indol-3-yl} ethyl) -2-amino-2-methylpropanamide; C 1-3 (C 1 -C 10) alkanoyloxy, C 1-3 (C 1 -C 6) alkoxy, phenyl, phenoxy, 2-furyl, (C 1-6) alkoxycarbonyl, (1-6) carbon channel) alkyl-O _m -S ···· ···· - may be substituted with 72 groups; or R ₄ and R _s together are - (CH _{2) r} L _a (CH _{2) s} - may form a group of the formula wherein L _a is -C (R ₂ ) ₂ -, -O-, -S (O) _m - or -N (R ₂ ) -, wherein r and s are independently 1-3 and R ₂ is as defined above; R ₆ is hydrogen or C 1-6 alkyl; A is a group of formula (a) or (b) wherein x and y are independently 0-3, R ₇ and R _7a are each independently hydrogen, C 1-6 alkyl, -OR ₂ , trifluoromethyl, phenyl, substituted C 1-6 alkyl, wherein the substituents are imidazolyl, phenyl, indolyl, p-hydroxyphenyl, -OR ₂ , 1-3 fluorine atoms, -S (O) _m R ₂ , -C (O) OR ₂ , C 3 -C 7 cycloalkyl, -N (R ₂ ) (R ₂ ), -C (O) N (R ₂ ) (R ₂ ) , obsession R ₇ and R _7a independently of one another may be linked to one or both of R ₄ and R ₅ to form an alkylene bridge between the terminal nitrogen atom and the alkyl moiety of R ₇ and R _7a which may contain from 1 to 5 carbon atoms; B, D, E and F are independently -C (R ₈ ) (R ₁₀ ) -, -O-, 73 C = O, -S (O) _in - or -NR ₉ - such that one or two of B, D, E, and F may be absent and thus a 5-, 6-, or 7-membered obtaining a ring; and with the proviso that B, D, E and F can be -C (R ₈ ) (R, _O ) - or C = O only if one of the remaining groups B, D, E and F is -0 -, -S (O) _m -, or -NR ₉ -, or B and D, or D and E together may be -N = CR ₁₀ - or -CR, _O = N-, or B and D, or D and E together may be -CR ₈ = CR, ₀ ; with the proviso that one of the other B and E or F is at the same time -O-, -S (O) _m - or -NR ₉ -; R ₈ and R ₁₀ are independently hydrogen, -R ₂ , -OR ₂ , - (CH ₂ ) _q -aryl, - (CH ₂ ) _q -C (O) OR ₂ , - (CH ₂ ) _q -C ( 0) O (CH ₂ ) _q -aryl or - (CH ₂ ) _q - (1H-tetrazol-5-yl) wherein the aryl group has 1 to 3 halogens, 1 or 2 C 1 -C 8 alkyl groups, 1 to 3 It may be substituted with 0R ₂ or 1 or 2 -C (O) OR ₂ ; R ₉ is -R ₂ , - (CH ₂ ) _q -aryl, -C (O) R ₂ , -C (O) (CH ₂ ) _q -aryl, -SO ₂ R ₂ , -SO ₂ (CH ₂ ) _{q is} -aryl, -C (O) N (R ₂ ) (R ₂ ), -C (O) N (R ₂ ) (CH ₂ ) _q -aryl, -C (O) OR ₂ , 1H-tetrazole-5 -yl, -SO ₃ H, -SO ₂ NHC = N, -SO ₂ N (R ₂ ) -aryl, -SO ₂ N (R ₂ ) (R ₂ ), and the moiety (CH ₂ ) _q optionally substituted with 1 or 2 C 1 -C 4 alkyl groups, and R ₂ and aryl optionally substituted with further substituents selected from 1-3 -OR _2a , -O (CH ₂ ) _q -aryl, 1- 2 -C (O) O R _2a , 1-2 -C (O) O (CH ₂ ) _q -aryl, 1-2 -C (O) N (R _2a ) (R _2a ), 1-2 - 74 -C (O) N (R _2a ) (CH ₂ ) _q -aryl, 1-5 halogen, 1-3C 1-4 alkyl, 1,2,4-triazolyl-, 1-H- tetrazol-5-yl, -C (O) NHSO ₂ R _2a , -S (O) _m R _2a , -C (O) NHSO ₂ (CH ₂ ) _q -aryl, -SO ₂ NHC = N, - SO ₂ NHC (O) R _2a , -SO ₂ NHC (O) (CH ₂ ) _q -aryl, -N (R ₂ ) C (O) N (R _2a ) (R _2a ), -N (R _2a ) C (O) N (R _2a ) (CH ₂ ) _q -aryl, -N (R _2a ) (R _2a ), -N (R _2a ) C (O) R _2a , -N (R _2a ) C (O) ) (CH ₂ ) _q -aryl, -OC (O) N (R _2a ) (R _2a ), -OC (O) N (R _2a ) (CH ₂ ) _q -aryl, -SO ₂ (CH ₂ ) _q CONH- (CH ₂ ) _w NHC (O) R wherein w is 2-6 and R _n is biotin, aryl, or aryl substituted with 1 or 2 OR ₂ , 1-2 halogen, azido or nitro groups; m is 0, 1 or 2; n is 1 or 2; q is optionally 0, 1, 2, 3 or 4; and G, Η, I and J are carbon, nitrogen, sulfur or oxygen, such that at least one of them is heteroatom and one of G, Η, I and J may be absent, resulting in a 5- or 6-membered heterocyclic aromatic ring develops. 1-9 halogen atoms, S (O) _m R _2a , 1-3 OR _2a or C (O) OR _2a , and the aryl groups are further substituted with phenyl, phenoxy, halophenyl, 1-3 C 6 alkyl groups, 1-3 halogens, 1-2 -OR ₂ groups, methylenedioxy groups, · · · · · · · · · · · · · · · · · · · · · · · ·· ·· · ··· - 71 -S (O) _m R ₂ groups, 1-2 _CF3 groups, -OCF _3, -NO _2, -N _{(R2) (R2),} -N (R ₂ ) C (O) R ₂ , -C (O) OR ₂ , -C (O) N (R ₂ ) (R ₂ ), -SO ₂ N (R ₂ ) (R ₂ ), - They may contain N (R ₂ ) S (O) ₂ -aryl and -N (R ₂ ) SO ₂ R ₂ ; R ₂ is hydrogen, C 1-6 alkyl, 1) NH ₂ , A combination for treating or preventing osteoporosis comprising a bisphosphonate and a growth hormone secretagogue. 2) pyridyl, 2) N- {1 (R) - [(1,2-Dihydro-1-methanecarbonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -2- (1 H- indol-3-yl} ethyl) -2-amino-2-methylpropanamide; 2) pyridyl, The combination of claim 1, wherein the bisphosphonate is a compound of formula (X), or a pharmaceutically acceptable salt thereof, wherein: R ¹ is (a) C 1-5 alkyl, unsubstituted or substituted with one of the following: 3) N- {1 (R) - [(1,2-Dihydro-1-benzenesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (1H -indol-3-yl) ethyl} -2-amino-2-methylpropanamide; C 3 -C 7 cycloalkyl, and where two C 1 -C 6 alkyl groups are present on an atom, they may optionally form a C 3 -C 8 ring, which may also contain oxygen, sulfur or NR _2a ; R _2a is hydrogen or C 1-6 alkyl; R _3a and R _3b are independently hydrogen, halogen, C 1-6 alkyl, -OR ₂ , cyano, -OCF ₃ , methylenedioxy, nitro, -S (O) _m R, -CF _3, or -C (O) OR ₂ , and when R _3a and R _{3b are in the} ortho position, they may together form a C 5 -C 8 aliphatic or aromatic ring optionally containing 1 or 2 heteroatoms selected from oxygen, sulfur and nitrogen; - 81 ·· ·· ···· ···· ·· • · · · · · · · · · · · · · • • · · · · · «·· ·· ·· * · R ₄ and R ₅ are each independently hydrogen, C 1-6 alkyl, C 1-6 alkyl having 1-5 halogens, 1-3 hydroxy, 3) pyrrodinyl, 3) N- {1 (R) - [(1,2-Dihydro-1-benzenesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (1H) -indol-3-yl) ethyl} -2-amino-2-methylpropanamide; • · • · - 76 4) Ν - {1 (R) - [(3,4-Dihydro-spiro [2 H -1-benzopyran-2,4'-piperidin] -1'-yl) carbonyl] -2- (1H -indol-3-yl) ethyl} -2-amino-2-methylpropanamide; C 3 -C 7 cycloalkyl, and where two C 1 -C 6 alkyl groups are present on an atom, they may optionally form a C 3 -C 8 ring, which may also contain oxygen, sulfur or NR _2a ; R _2a is hydrogen or C 1-6 alkyl; R _3a and R _3b are independently hydrogen, halogen, C 1-6 alkyl, -OR ₂ , cyano, -OCF ₃ , methylenedioxy, nitro, -S (O) _m R, -CF _3, or -C (O) OR ₂ , and when R _3a and R _{3b are in the} ortho position, they may together form a C 5 -C 8 aliphatic or aromatic ring optionally containing 1 or 2 heteroatoms selected from oxygen, sulfur and nitrogen; R ₄ and R _s are independently hydrogen, C1-6 alkyl, C1-6 alkyl, 1-5 halogen atoms, 1-3 hydroxy groups, The combination according to claim 1, wherein the bis'phonate is alendronic acid, etidronic acid, clodronic acid, pamidronic acid, tiludronic acid, rizedronic acid, 6-amino-1-hydroxyhexylidene bisphosphonic acid and 1-hydroxy-3- (methylpentylamino). propylidene ·· ·· ·· ···· ········ · · · · • "·· · · · · · · · · · • · · ···· ··" ·· - 70-bisphosphonic acid or a pharmaceutically acceptable salt thereof. 3) pyrrodinyl, »· ·· ···· ···· ··" 4 * * · · · · »· · · · · · · · · · ·· • ·« »* ·« · ·· ·· - 86 i 4) N- {1 (R) - [(3,4-Dihydro-spiro [2H-1-benzopyran-2,4'-piperidin] -1'-yl) carbonyl] -2- (1H-indole- 3-yl} ethyl) -2-amino-2-methylpropanamide; 4) NR ³ R ⁴ (b) NR ⁵ , (c) SR ⁶ or d) Cl; R ² is H, OH or Cl; R ³ is H or C 1-4 alkyl, R ⁴ is C 1 -C 4 alkyl, R ⁵ is C 1 -C 10 alkyl and - 79 R ⁶ is aryl. The combination of claim 3, wherein the bisphosphonate is alendronic acid or a pharmaceutically acceptable salt thereof. 4) NR ³ R ⁴ (b) NR ⁵ , (c) SR ⁶ or d) Cl; R ² is H, OH or Cl; R is H or C 1-4 alkyl, R ⁴ is C 1 -C 4 alkyl, R ⁵ is C 1 -C 10 alkyl and R ⁶ is aryl. i. 5) N- {1 (R) - [(2-acetyl-1,2,3,4-tetrahydro-spiro [isoquinolin-4,4'-piperidin] -1'-yl) carbonyl] -2- ( indol-3-yl} ethyl) -2-amino-2-methylpropanamide; 5) N- {1 (R) - [(2-acetyl-1,2,3,4-tetrahydro-spiro [isoquinolin-4,4'-piperidin] -1'-yl) carbonyl] -2- ( indol-3-yl} ethyl) -2-amino-2-methylpropanamide; A combination according to claim 1, which comprises as a growth hormone secretagogue a compound of formula (I) or (II), or a pharmaceutically acceptable salt thereof, or a single diastereomeric form thereof, wherein: R.! is C 1-10 alkyl, aryl, (C 1-6) alkylaryl, (C 1-6) alkyl (C 3-7) cycloalkyl, (C 1-5) alkyl-K-, (C1-C5) alkyl, (C1-C5) alkyl-K (C5-C5) alkylaryl, (C1-C5) alkyl-K (C5-C5) alkyl- C 3 -C 7) cycloalkyl, where K is O, S (O) _m , N (R ₂ ) C (O), C (O) N (R ₂ ), OC (O), C (O) O or -CR ₂ = CR ₂ - or - C = C and aryl have the meanings given below and R ₂ and alkyl are further substituents 6) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenyl) methyloxy) ethyl} -2-amino-2-methylpropanamide; 6) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenyl- methyloxy) ethyl} -2-amino-2-methyl-propánainid; A combination according to claim 1, which comprises as a growth hormone secretagogue a compound of formula (V), or a pharmaceutically acceptable salt or individual diastereomer thereof, wherein: R j is a group of the formula (c), (d), (e), (f), (g), (h), (i), (j), (k) or (1); RJA ^e l j Rs are hydrogen or fluoro; • · · · • · · - 75 D is -Ο-, -S-, -S (O) _m -, N (R ₂ ), NSO ₂ (R ₂ ), NSO ₂ (CH ₂ ) _t -aryl, NC (O) (R _2). ), NSO ₂ (CH ₂ ) _q OH, NSO ₂ (CH ₂ ) _q COOR ₂ , NSO ₂ (CH ₂ ) _q C (O) -N (R ₂ ) (R ₂ ), N-SO ₂ (CH ₂₎ ) _q C (O) -N (R ₂ ) (CH ₂ ) _w OH, or a group of formula (m), (n) or (o), and aryl represents a phenyl or pyridyl group and the phenyl group has 1 to 2 halogens. may be substituted; R ₂ is hydrogen or C 1-4 alkyl; m is 1 or 2; t is 0, 1 or 2; q is 1, 2 or 3; w is 2, 3, 4, 5 or 6. 7) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenyl) methyloxy) ethyl} -2-amino-2-methylpropanamide mesylate salt; 7) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) -carbonyl] -2- (phenyl- methyloxy) ethyl} -2-amino-2-methylpropanamide mesylate salt; The combination of claim 1, wherein the growth hormone secretagogue is a compound selected from the group consisting of or a pharmaceutically acceptable salt thereof: 8) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (2 ', 6'-difluoro-phenylmethyl) ethyl} -2-amino-2-methylpropanamide; The combination of claim 1, wherein the bisphosphonate is alendronic acid or pamidronic acid, or a pharmaceutically acceptable salt thereof, and a growth hormone secretagogue. N- {l (R) - 2- (phenyl-methyl-oxy [(l, 2-dihydro-l-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -r-yl) carbonyl] } ethyl) -2-amino-2-methyl-propanamide mesylate salt; obsession N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl} -2 -ainino-2-methylpropanamide, or a pharmaceutically acceptable salt thereof. • · · · • · · The combination of claim 1, wherein the bisphosphonate is alendronic acid or a pharmaceutically acceptable salt thereof and a growth hormone secretagogue. N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indo-3,4,4'-piperidin] -1'-yl) carbonyl] -2- (phenyl- methyloxy) ethyl} -2-amino-2-methylpropanamide, or a pharmaceutically acceptable salt thereof. 8) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -2- (2 ') 6'-Difluorophenylmethyloxy) ethyl} -2-amino-2-methylpropanamide; 9) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-5-fluoro-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2 - (phenyl-methyl) ethyl} -2-amino-2-methylpropanamide; 9 9 - 82 R ₇ and R _{7; 1} independently of one another R ₄ and R _5, or both, may form an alkylene bridge between the terminal nitrogen atom and the alkyl moiety of R ₇ and R _7a groups which may have from 1 to 5 carbon atoms; B, D, E and F are each independently -C (R ₈ ) (R ₁₀ ) -, -O-, C = O, -S (O) _m - or -NR ₉ - such that one or two of B, D, E and F may be absent to give a 5-, 6- or 7-membered ring ; and provided that B, D, E, and F are can only be -C _{(R8) (R10)} - or C = O, where B, D, E and F are, however, one of the remaining groups -O- , -S (O) _m - or -NR ₉ -, or B and D, or D and E together represent -N = CR, _O - or -CR ₁₀ = N-, or B and D, or D and E together represent -CR ₈ = CR ₁₀ ; with the proviso that one of the other B and E or F is at the same time -O-, -S (O) _m - or -NR ₉ -; R ₈ and R ₁₀ are independently hydrogen, -R ₂ , -OR ₂ , - (CH ₂ ) _q -aryl, - (CH ₂ ) _q -C (O) OR ₂ , - (CH ₂ ) _q -C ( O) O (CH ₂ ) _q -aryl or - (CH ₂ ) _q - (1 H -tetrazol-5-yl) wherein the aryl group has 1 to 3 halogens, 1 or 2 C 1 -C 8 alkyl groups, 1 to 3 -OR _It may be substituted with ₂ groups or 1 or 2 -C (O) OR ₂ groups; R ₉ is -R ₂ , - (CH ₂ ) _q -aryl, -C (O) R ₂ , -C (O) (CH ₂ ) _q -aryl, -SO ₂ R ₂ , -SO ₂ (CH ₂ ) _q -aryl, -C (O) N (R ₂ ) (R ₂ ), -C (O) N (R ₂ ) (CH ₂ ) _q -aryl, -C (O) OR ₂ , 1H-tetrazole-5 -il group, · · · ··· · · · · · · · · · · · - 83 -SO ₃ H, -SOpNHCsN. -SO ₂ N (R ₂ ) -aryl, -SO ₂ N (R ₂ ) (R ₂ ), and (CH ₂ ) _q may be optionally substituted with 1 or 2 C 1 -C 4 alkyl groups, and R ₂ and aryl optionally substituted with further substituents selected from the group consisting of 1-3 -OR _2a , -O (CH ₂ ) _q -aryl, 1-2 -C (O) OR _2a , 1-2 -C (O) O (CH ₂ ) _q -aryl, 1-2-C (O) N (R _2a ) (R _2a ), 1-2 -C (O) N (R _2a ) (CH ₂ ) _q -aryl, 1-5 halogen, 1-3 C 1-4 alkyl, 1,2,4-triazolyl, 1-H-tetrazole-5- yl, -C (O) NHSO ₂ R _2a , -S (O) _m R _2a , -C (O) NHSO ₂ (CH ₂ ) _q -aryl, -SO ₂ NHC = N, -SO ₂ NHC ( O) R _2a , -SO ₂ NHC (O) (CH ₂ ) _q -aryl, -N (R ₂ ) C (O) N (R _2a ) (R _2a ), -N (R _2a ) C (O) N (R _2a ) (CH ₂ ) _q -aryl, -N (R _2a ) (R _2a ), -N (R _2a ) C (O) R _2a , -N (R _2a ) C (O) (CH ₂₎ ) _q -aryl, -OC (O) N (R _2a ) (R _2a ), -OC (O) N (R _2a ) (CH ₂ ) _q -aryl, -SO ₂ (CH ₂ ) _q CONH- (CH _{2) w} NHC (O) Ra, _I5 wherein w is 2-6, and R _u is biotin, aryl, or 1 or 2 OR ₂ groups, 1-2 halogen, azido or nitro szubsztitutált aryl; m is 0, 1 or 2; n is 1 or 2; q is optionally 0, 1, 2, 3 or 4; and G, Η, I and J are carbon, nitrogen, sulfur or oxygen, such that at least one of them is heteroatom and one of G, Η, I and J may be absent, resulting in a 5- or 6-membered heterocyclic aromatic ring develops. - 84 t 9) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-5-fluoro-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -2 - (phenyl-methyl) ethyl} -2-amino-2-methylpropanamide; 10) Ν- {1 (S) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyl) thio) ethyl} -2-amino-2-methylpropanamide; A pharmaceutical composition for treating or preventing osteoporosis comprising a combination of a bisphosphonate and a growth hormone secretagogue and an inert carrier. 10) N- {1 (S) - [(1,2-dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) -carbonyl] -2- (phenyl- methylthio) ethyl} -2-amino-2-methylpropanamide; 11. The composition of claim 10, wherein the bisphosphonate is a compound of formula (X), or a pharmaceutically acceptable salt thereof, wherein: R ¹ is (a) C 1-5 alkyl, unsubstituted or substituted with one of the following: 11) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indol-3,4'-piperidin] -1'-yl) carbonyl] -3-phenyl-propyl } -2-amino-2-methylpropanamide; 12) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-cyclohexyl-propyl} -2-amino-2-methylpropanamide; The composition of claim 10, wherein the bisphonate is alendronic acid, etidronic acid, clodronic acid, pamidronic acid, tiludronic acid, rizedronic acid, 6-amino-1-hydroxyhexylidene bisphosphonic acid and 1-hydroxy-3- (methylpentylamino) propylidene bisphosphonic acid. , or a pharmaceutically acceptable salt thereof. 12) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -3-cyclohexyl-propyl } -2-amino-2-methylpropanamide; • · - 77 13) Ν- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -4-phenyl butyl} -2-amino-2-methylpropanamide; 13) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -4-phenyl-butyl} -2-amino-2-methylpropanamide; The composition of claim 12, wherein the bisphosphonate is alendronic acid or a pharmaceutically acceptable salt thereof. 14) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (5- fluoro-lH-indol-3-yl} ethyl) -2-amino-2-methylpropanamide; 14. A composition according to claim 10, which comprises as a growth hormone secretagogue a compound of formula (I) or (II), or a pharmaceutically acceptable salt thereof, or a single diastereomeric form thereof, wherein: R 1 is C 1 -C 10 alkyl, aryl, (C 1 -C 6) alkyl-aryl, (C 1 -C 6) alkyl (C 3 -C 7) cycloalkyl, (C 1 -C 5) alkyl-K-. (C 1 -C 5) alkyl, (C 1 -C 5) alkyl-K (C 0 -C 5) alkylaryl, (C 1 -C 5) alkyl-K (C 5 -C 5) alkyl- (C3-C7) cycloalkyl, wherein K is O, S (O) _m , N (R ₂ ) C (O), C (O) N (R ₂ ), OC (O), C (O) O or -CR ₂ = CR ₂ - or - C = C and aryl have the meanings given below and R ₂ and alkyl are further substituents 14) Ν - {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-1-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (5-Fluoro-1H-indol-3-yl) -ethyl} -2-amino-2-methyl-propanamide; 15) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-5-fluoro-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2 - (5-Fluoro-1H-indol-3-yl) -ethyl} -2-amino-2-methyl-propanamide; 15. A composition according to claim 10, which comprises as a growth hormone secretagogue a compound of formula (V), or a pharmaceutically acceptable salt or individual diastereomer thereof, wherein: R 1 is a group of formula (c), (d), (e), (f), (g), (h), (i), (j), (k) or (1); R _3a is hydrogen or fluorine; D is -O-, -S-, -S (O) _m -, N (R ₂ ), NSO ₂ (R ₂ ), NSO ₂ (CH ₂ ) _t -aryl, NC (O) (R ₂ ), NSO ₂ (CH ₂ ) _q OH, NSO ₂ (CH ₂ ) _q COOR ₂ , NSO ₂ (CH ₂ ) _q C (O) -N (R ₂ ) (R ₂ ), N-SO ₂ (CH ₂ ) _q C (O) -N (R ₂ ) (CH ₂ ) _w OH, or (m), (n) or (o), and aryl represents phenyl or pyridyl, and the phenyl may be substituted with 1 to 2 halogens. ; R ₂ is hydrogen or C 1-4 alkyl; m is 1 or 2; t is 0, 1 or 2; q is 1, 2 or 3; w is 2, 3, 4, 5 or 6. 15) N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-5-fluoro-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (5-fluoro-lH-indol-3-yl} ethyl) -2-amino-2-methylpropanamide; 16) N- {1 (R) - [(1,2-Dihydro-1- (2-ethoxycarbonyl) methylsulfonyl-spiro [3H-indole-3,4'-piperidin-1'-yl) carbonyl] ] -2- (1H-indol-3-yl) -ethyl} -2-amino-2-methyl-propanamide; 16. The composition of claim 10, wherein said growth hormone secretagogue is selected from the group consisting of or a pharmaceutically acceptable salt thereof. 16) N- {1 (R) - [(1,2-Dihydro-1- (2-ethoxycarbonyl) methylsulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) - carbonyl] -2- (1H-indol-3-yl) ethyl} -2-amino-2-methylpropanamide; The composition of claim 10, wherein the bisphosphonate is alendronic acid or pamidronic acid, or a pharmaceutically acceptable salt thereof, and a growth hormone secretagogue. N- {1 (R) - [(1,2-Dihydro-1-methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyl) butyldimethylsilyloxy) ethyl} -2-amino-2-methyl-propanamide mesylate salt; obsession - 87 ⁷ ί Ν- {1 (R) - [(1,2-dihydro-1 -methanesulfonyl-spiro [3H-indole-3,4'-piperidin] -1' -jl) carbonyl] -3-phenyl -propyl} -2-amino-2-methylpropanamide, or a pharmaceutically acceptable salt thereof. 17) N- {1 (R) - [(1,2-Dihydro-1,1-dioxo-spiro [3H-benzothiophene-3,4'-piperidin] -1'-yl) carbonyl] -2- ( phenyl-methyl) ethyl} -2-amino-2-methylpropanamide. 17) N- {1 (R) - [(1,2-Dihydro-1,1-dioxo-spiro [3H-benzothiophene-3,4'-piperidin] -1'-yl) carbonyl] -2- ( phenyl-methyl) ethyl} -2-amino-2-methylpropanamide. 18. The composition of claim 10, wherein the bisphosphonate is alendronic acid or a pharmaceutically acceptable salt thereof and the growth hormone secretagogue is N- {1 (R) - [(1,2-dihydro-1-methanesulfonyl-spiro [3H-indole] 3,4'-piperidin] -1'-yl) carbonyl] -2- (phenylmethyloxy) ethyl} -2-amino-2-methylpropanamide, or a pharmaceutically acceptable salt thereof.