CZ607390A3 - Octapeptide analogs of somatostatin, having threonine in position six - Google Patents

Abstract

A compound of formula wherein each of A 1 and A2 is independently H, C1-6alkyl, C7. lophenylalkyl, R 1CO, wherein R 1 is C 1-20 alkyl, C 3-20 alkenyl, C3-20alkynyl, phenyl, naphthyl or C7-phenylalkyl, or R2OCO, wherein R 2 is C 10-10 alkyl or C 7-10 phenylalkyl with halo provided that when one of the substituents A! or A2 is RiCO or R2OCO, the other must be H, each X1 and X2 independently of one another are H, F, Cl, Br, OH, CH 3 or CF 3 with the proviso that at least one of X1 and X2 must be H, A3 is Phe or Tyr and A) is OH, NH 2 or NH-R 3 wherein R 3 is saturated aliphatic C 14 alkyl, or a pharmaceutically acceptable salt thereof compounds; a therapeutic composition comprising the same the compounds and the method of using the compounds.

Description

Technical field

The present invention relates to therapeutically effective peptides.

BACKGROUND OF THE INVENTION

A considerable number of somato-tatato analogs are described in the literature

Veins exhibiting growth Mi release inhibiting activity

including analogues comprising at least fourteen amino acids of natural origin. For example, Coy et al. U.S. Patent No. 4,485,101 discloses dodecapeptides having an amino-terminal acetyl group, a carboxy-terminal amino group, D-Trp at position 6 and p-Cl-Phe at position 4. (Each amino acid is designated by its standard three letter abbreviation herein, stereoisomeric designation of each amino acid) is L unless otherwise stated).

SUMMARY OF THE INVENTION

The present invention provides compounds of the following formula

X,

CH-C-Cys-Ao-D-Trp-Lys-Thr-C.ys- / 3 Nal-A-2

-2where each and Ag, independently of the other, is H, C 1-4 alkyl,

IQfenylalkyl Cy, R₁ CO (where R '^ is' ^C 3-2O ^{alkenyl> C} 3-2O ^{alkynyl' -phenyl> naphth-yl,} or C7_iQfenylalkyl) RgOCO or (wherein Rg is C ^ _ ^ ^ q alkyl or Ογ_ θί en) (1) with the proviso that when one of the substituents A 4 or Ag is R 6 CO or R 8 COCO, the other of the substituents H must be H;

each X 1 and X g, independently of the other, is H, F, Cl, Br, OH, CH 2 or CF 1, provided that at least one of X 1 and X g must be H;

ky is Phe or T.yr; and

A ^ is OH, or NHG NH-R (wherein R represents a saturated aliphatic ^C 8 ^alk-yi >>

or a pharmaceutically acceptable salt thereof. Amino acids of natural origin are identified by their commonly used three-letter symbols; if the D-stereoisomer of the amino acid (other than NaI) is not labeled, the L-form is used. "/ 3 Nal" means D- or L-naphthylalanine unless a D- or L-stereoisomer is specified.

In a preferred embodiment, each of A 1 and Ag independently of the other, is H or saturated aliphatic C 1-6 alkyl; each X 1 and X g independently of the other, is H, F, Cl or OH, with the proviso that at least one of X y and X g must be H; and R 6 is saturated aliphatic C 1-4 alkyl;

more preferably, the compound has the following formula:

D -P h e -Cy s -P h e -D -T r p-Ly s -T hr -Cy s-fb Na 1 -NH, or

D-Phe-Cys-Tyr-L-Trp-Lys-Thr-Cys-2 Nal-NHg

In another aspect, the present invention relates to compounds of the formula:

D-β-Nal-Cys -Phe-D-Trp-Lys -Thr-Cys -Thr-NH

L - / Nal-Cys -Tyr -D -Tr p-Lys -Tr-Cys -Tr-Nr or a pharmaceutically acceptable salt thereof.

The present invention also relates to a combination of one of the above compounds and a pharmaceutically acceptable carrier subv

Stances in a therapeutic composition capable of inhibiting the release of growth hormone ("GH"), epidermal growth factor, insulin, glukayone, pancreatic exocrine secretions or substance P and preferred GH.

In preferred embodiments, the formulation is in the form of pills, tablets, capsules or liquid preparations for oral administration; a cream, gel, lotion, spray or ointment for application to the skin of a patient; a liquid which can be administered nasally as drops or drops; or liquids suitable for entravenous, subcutaneous, parenteral or intraperitoneal administration. The therapeutic composition may also be in the form of an oil emulsion or or dispersion together with a lipophilic salt such as a pamoic acid salt, or in the form of a biodegradable, sustained release formulation for subcutaneous or intra-muscular administration. A zero release order is desirable for maximum efficiency. This order may be obtained using an implant or an external pump for administration of the therapeutic agent.

The compounds of the invention exhibit a wide range of biological activities due to their antisecretory and antiproliferative properties. The compounds inhibit the secretion of some endocrine hormones, including insulin, glucagon, and especially growth hormone (GH). The compounds of the invention also suppress pancreatic and gastric exocrine secretions and suppress or modulate the release of certain neurotransmitters including substance P and acetylcholine.

Somatostatin analogs may act to multiply the tumor cell by preventing the release of mitotic factors (such as insulin-like growth factor 1 (IGF-1), gastrin releasing peptides, etc.) and may interfere with an intracellular transduction mechanism such as enidermal growth factor (EGF) - inducing cell proliferation.

The aromatic lipophilic N-terminal end may provide long-lasting in vivo activity.

Other features and advantages of the present invention will become apparent from the following description of the preferred embodiments and from the claims.

The compounds of the invention, which are peptide analogs of somatostatin, have the above formula.

The compounds may be in the form of pharmaceutically acceptable salts or complexes. As used herein, the term "pharmaceutically acceptable salts or complexes" refers to salts or complexes that retain the biological activity of the parent compounds and do not produce any undesirable toxicological effects. Examples of such salts are a) acid addition salts which are inorganic acids (e.g. hydrochloric acid, hydrobromic, sulfuric, phosphoric, nitric and the like), and salts formed with organic acids such as acetic acid,

-5-oxalic, tartaric, succinic, malic, ascorbic, benzoic, tannin, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acids, naphthalenedisulfonic acids, and polygalacturonic acid;

(b) base addition salts formed with polyvalent metal cations such as zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, nickel, cadmium and similar metals, or with organic cations derived from Ν, Ν-dibene » z.ethylethylenediamine or ethylenediamine; or

(c) combinations (a) and (b): eg, zinc tannin salt or similar salts.

Examples

The synthesis of octapeptides is given below. Other compounds of the invention may be prepared using suitable modifications, which will be apparent to those skilled in the art, from the following synthetic methods.

First stage in preparation

D-Phe-Cys-Phe-D-Trp-Lys-Val-Cys-Nal-NH2 is the preparation of the intermediate t-butyloxycarbonyl-D-Phe-S-methylbenzyl-Cysc

Phe-D-Trp-N-b Enoxycarbonyl-Lys-Thr-S-methylb-1-Cys-Nal-N 1 -benzhydrylamine resin as follows.

Methyl-benzhydrylamine-polystyrene resin (Advanced Chem-Tech, Inc) in chloride ion form is placed in a reaction vessel of a Beckman 99OB peptide synthesizer programmed to perform the following reaction cycles:

(a) methylene chloride; b) 33% trifluoroacetic acid in methylene chloride (2 times every 2 and 25 minutes); c) methylene chloride; (d) ethanol; e) methylene chloride; f) 10% triethylamine in chloroform.

The 6-neutralized resin was stirred with Boc- / 3Nal and diisopropylcarhodiimide (1.5 mmol each) in methylene chloride for 1 hour, and then the resulting amino acid resin was cycled through steps a) to f) of the above wash program. The following amino acids (1.5 mmol) are then coupled in the same manner: Boc-S-methylbenzyl-Cys, Boc-Thr, Boc-N-benzyloxycarbonyl-lysine, Boc-D-Trp, Boc-Phe, Boc-Smethylbenzyl- Cys, Boc-D-Phe.

The resin was washed and dried and then mixed with anisole (4 mL) and anhydrous hydrogen fluoride (36 mL) at 0 ° C, then stirred for 45 min. Thioanisole, trifluoroacetic acid and trifluoromethanesulfonic acid 1:90 can also be used: 9 for 6 hours. Excess hydrogen fluoride was evaporated rapidly with a stream of dry nitrogen and the free peptide precipitated and washed with ether. The crude peptide is then dissolved in 800 ml of 90% acetic acid, to which is added 10% acetic acid. in methanol until a permanent brown color is obtained. The solution was then stirred for 1 hour before the solvent was removed in vacuo. The resulting oil was dissolved in a minimum volume of 50% acetic acid and eluted on a column (2.5 x 100 mm) of Sephadex G-25. The fractions containing the main component by UV absorption and thin layer chromatography are then combined, evaporated to a small volume and loaded onto a 2.5 cm x 50 cm Vydac octadecylsilane (1015 µM) column.

The column was eluted with a linear gradient of 10 to 50% acetonitrile in 0.1% trifluoroacetic acid in water. The fractions are analyzed by thin layer chromatography and high pressure liquid chromatography, combined to the maximum purity and, if desired, various salts are prepared, e.g. acetate or phosphate. Repeated lyophilization of the solution from water gave 120 mg of the product as a white, fluffy powder.

The product was found to be homogeneous by the results of high pressure liquid chromatography and thin layer chromatography

-7 May. Analysis of the amino acids in the acid hydrolyzate shows that it is an octadecapeptide.

Compounds of the invention having the formulas

may be prepared by methods analogous to those described above.

Usability

When administered to mammals, particularly humans (eg, orally, topically, intravenously, sustained-release biodegradable form, nasally or suppository), these compounds can effectively inhibit the secretion of various hormones and trophic factors. They can be used to suppress certain endocrine secretions such as GH, insulin, glucagon and prolactin in the treatment of, for example, acromegaly, endocrine tumors such as carcinoids, vipomas, insulinomas and glucagon hormones, or diabetes and diabetes-induced diseases, nephropathy, fatigue and diabetes type 2. The compounds may also be used to suppress exocrine secretions in the pancreas, stomach and intestines in the treatment of, for example, pancreatitis, fistula, bleeding ulcers and diarrhea associated with diseases such as AIDS or cholera. Disorders associated with autocrine or paracrine secretion of trophic factors such as IGF-1 (as well as some endocrine factors) that can be treated by administration of these compounds include breast, prostate and lung cancers (both small and non-small cell epidermoids) as well as hepatomas, neuroblastomas,

-8 adenocarcinomas of the intestine and pancreas (ductal type), chondrosarcomas and melanomas, as well as atherosclerosis associated with vascular transplants and restenosis of subsequent angioplasty.

The compounds of the invention are also useful for suppressing mediator of neurogenic inflammation (eg substance P or tachine kinins) and can be used to treat pathological conditions such as rheumatoid arthritis, psoriasis, skin inflammations such as sunburn, eczema or other sources of itching. ; and allergies, including asthma. The compounds may also act as neuromodulators in the central nervous system and may be used in the treatment of Alzheimer's disease and other forms of dementia, pain (such as spinal analgesia) and headache. Furthermore, the compounds of the invention may provide cell protection in disorders related to the intestinal blood flow, including cirrhosis, oesophageal varices, and in some cases, fungal poisoning.

The compounds may be administered to mammals, e.g., humans, at a dose of 0.01 to 50 mg / kg / day, preferably 0.1 to 5 mg / kg / day.

Other embodiments are within the scope of the following claims.

Claims (13)

PATENT CLAIMS Compound of formula CH, and 6 o « ^{0 0} f-5 N-CH-C-Cys-N-D-Trp-Lys-Thr-Gys-Nal-A-O where each of A 1 and A 3, independently of the other, is H, alkyl of 1 to 12 carbon atoms, phenylalkyl C 7 -C 10, R 4 CO, wherein R 4 is alkyl of 1 to 20 carbon atoms, alkynyl of 3 to 20 carbon atoms, phenyl, naphthyl or phenylalkyl of 7 to 10 carbon atoms, or R 2OCO, wherein R 2 is alkyl of 1 to 10 carbon atoms or phenylalkyl of 7 to 10 carbon atoms, with the proviso that when one of A 1 or Ag is R 1 CO or R 2 OCO, the other H, each substituent and independently on the other hand, H, F, Cl, Br, OH, CH 2 or CF 3, provided that at least one of X ₁ and X ₂ is H, is Phe or Tyr and A 1 is OH, NH 3 or NH-R 6, wherein R 1 is saturated aliphatic alkyl of 1 to 8 carbon atoms, or a pharmaceutically acceptable salt thereof. -lo2. A compound according to claim 1, wherein each of the substituents Q independently of the other represents H, or saturated aliphatic alkyl group having 1 to 3 carbon atoms, each substituent XYa ^{X2 than} independently, an another represent H, F, Cl or OH, with the proviso that at least one of X 1 and X 2 must be H and is C 1 -C 3 aliphatic alkyl, or a pharmaceutically acceptable salt thereof. The compound of claim 2 having the formula D-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-N-Nal-N-4 or a pharmaceutically acceptable salt thereof. The compound of claim 2 having the formula D-Phe-Cys -Tyr-D-Trp-Lys-Thr-Cys-N-Nal-NH 4 or a pharmaceutically acceptable salt thereof. 5. Compound of formula D- / 3 Nal-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-NH ₂ or D-Na 1 -Cys -Tyr-D-Trp-Lys-Thr-Cys-T hr -N ^ or a pharmaceutically acceptable salt thereof -116. A therapeutic composition capable of inhibiting the release of GH, epidermal growth factor, insulin, glucagon, prolactin, exocrine secretions from the pancreas, stomach or intestine, tachycining and substance P, comprising a therapeutically effective amount of the compound of claim 1 or 5, together with a pharmaceutically acceptable carrier of the substance. 7. A therapeutic composition according to claim 6 which is capable of inhibiting the release of GH. A therapeutic composition according to claim 6 which is in the form of a liquid, pills, tablets or capsules for oral administration of the composition to human patients in need thereof. 9. A therapeutic composition according to claim 6 which is in the form of a cream, gel, lotion, spray or ointment for application to the skin of a human patient in need thereof. A therapeutic composition according to claim 6 which is in the form of a liquid suitable for nasal administration as drops or spray to human patients if desired. 11. A therapeutic composition according to claim 6 which is in the form of a liquid for intravenous, subcutaneous, parenteral or intraperitoneal administration to human patients in need thereof. 12. A therapeutic composition according to claim 6, wherein the therapeutic composition is in the form of a biodegradable sustained release formulation for intranumicular administration to human patients in need thereof. -12 * A therapeutic composition according to claim 6, characterized in that it comprises a lipophilic salt and is suitable for administration as an oil emulsion or dispersion to human patients if desired. 14. A method of treating mammals in need of reduction of GN, epidermal growth factor, insulin, glucagon, prolactin, exocrine secretions from the pancreas, stomach or intestine, tachycining and substance P, comprising administering a therapeutically effective amount of a compound of claim 1 or claim 5 to said mammal.