Kahalalide f derivatives

FIELD: chemistry.

SUBSTANCE: invention relates to novel anti-tumour compounds which are kahalalide F derivatives, pharmaceutical compositions containing said derivatives and their use in preparing an anti-tumour medicinal agent.

EFFECT: obtaining novel anti-tumour compounds.

4 cl, 9 tbl, 17 ex

 

The present invention relates to new kahalalide antitumor compounds, and in particular analogs kahalalide F containing their pharmaceutical compositions and to their use as antitumor, antiviral, antifungal agents in the treatment of psoriasis.

Kahalalide compounds are peptides isolated from Hawaiian herbivorous marine species of mollusk, Elysia rufescens, and their food, green algae Bryopsis sp. Kahalalide F described Hamann, M. and others, J. Am. Chem. Soc., 1993, 115, 5825-5826.

Kahalalide A-G described Hamann, M. and others, J. Org. Chem., 1996, 61, 6594-6600: "Kahalalides: bioactive peptides from a marine mollusk Elysia rufescens and its algal diet Bryopsis sp.".

Kahalalide N and J described Scheuer P.J., and others, J. Nat. Prod., 1997, 60, 562-567: "Two acyclic kahalalides from the sacoglossan mollusk Elysia rufescens".

Kahalalide About described Scheuer P.J., and others, J. Nat. Prod., 2000, 63(1), 152-4: "A new depsipeptide from the sacoglossan mollusk Elysia ornata and the green alga Bryopsis species".

In relation kahalalide To see Kan, Y. and others, J. Nat. Prod., 1999, 62(8), 1169-72: "Kahalalide K: A new cyclic depsipeptide from the hawaiian green alga Bryopsis species".

In respect of related messages see also Goetz and others, Tetrahedron, 1999, 55, 7739-7746: "The absolute stereochemistry of Kahalalide F"; Albericio F. and others, Tetrahedron Letters, 2000, 41, 9765-9769: “Kahalalide B. Synthesis of a natural cyclodepsipeptide”; Becerro, etc., J. Chem. Ecol., 2001, 27(11), 2287-99: “Chemical defenses of the sarcoglossan molluskElysia rufescensand its host Algabryopsis sp.”.

From kahalalide compounds kahalalide F is the most promising because of its antitumor the activity. He has a complex structure consisting of six amino acids as cyclic part and ekzoticheskuyu chain of seven amino acids with the terminal group of the fatty acid. Originally kahalalide F corresponds to the structure (I):

In WO 04035613 described 4(S)-methylhexane compound of the following formula (II). WO 04035613 fully included in this context by reference.

Activity kahalalide F againstin vitrocell cultures of human lung carcinoma A-549 and carcinoma of the colon of human HT-29 described in EP 610078. Kahalalide F also exhibits antiviral and antifungal properties, and is also suitable for the treatment of psoriasis.

In WO 0236145 describes pharmaceutical compositions containing kahalalide F, new applications of this compound in cancer therapy, and its contents are fully incorporated into this context by reference.

In WO 0333012 describes the clinical application of kahalalide compounds in Oncology, and its contents are fully incorporated into this context by reference.

Synthesis and cytotoxic activity of natural and synthetic kahalalide compounds described in WO 0158934, which is fully included in this context as a reference. In WO 0158934 describes the synthesis kahalalide F and also compounds of similar structure in which the terminal t the universi fatty acids are replaced with other fatty acids.

However, there is a need to further anticancer compounds, in particular, further kahalalide compounds with improved properties.

Summary of the invention

Received similar kahalalide compounds with improved biological activity.

The present invention relates to compounds of formula (1):

where one or more amino acids in the cyclic or ekzoticheskoy part replaced by other natural or unnatural amino acids protected by organic groups, or deleted. The present invention also relates to compounds of formula (1), where the aliphatic acid end group is replaced by other acyl groups, or deleted.

The present invention also relates to pharmaceutical compositions containing the above compound and a pharmaceutically acceptable carrier, solvent or diluent.

The present invention further relates to a method of treating any mammal, especially human, cancer, viral infection, fungal infection or psoriasis, which includes the introduction of the affected individual a therapeutically effective amount of a compound as described above.

The present invention particularly can be used for the treatment of PA is antov with intractable cancers, which are not amenable to the effects of other treatments. In particular, compositions according to this invention can be applied after it has been tested and did not produce results different chemotherapy.

The present invention particularly relates to the treatment of patients affected by prostate cancer, breast cancer, hepatocellular cancer, melanoma, cancer of the colon and rectum, kidney cancer, ovarian cancer, NSCL cancer, epithelial cancer, pancreatic cancer and tumors, which sverkhekspressiya the oncogene Her2/neu.

In another aspect, the present invention relates to the use of compounds as described above, to obtain drugs. In the preferred embodiment the drug is a drug for the treatment of cancer, psoriasis, viral infection or fungal infection.

This invention additionally provides a pharmaceutical kit containing a separate container filled with a pharmaceutical composition comprising the above compound and reducing agent. Also provides methods of recovery.

The authors identified analogues kahalalide F, which show a significant improvement in activity relative to kahalalide F.

The present invention relates to compounds of formula (1):

where one or more ekzoticheskih or cyclic amino acids replaced by other natural or unnatural amino acids protected by organic groups, or deleted. The present invention also relates to compounds of formula (1), where the aliphatic methylhexaneamine group substituted by other acyl groups, or deleted.

Ekzoticheskie amino acids

Preferred compounds according to this invention correspond to the formula (1), where one or more amino acids ekzoticheskoy chain replaced by other natural or unnatural amino acids protected by organic groups, or removed.

Especially preferred compounds include those with substitution of 1, 2 or 3 amino acids in ekzoticheskoy chain and any remote 1, 2, 3, 4, 5 or 6 amino acids in ekzoticheskoy chain.

Especially preferred are such compounds of formula (1), where one ekzoticheskaya amino acid substituted by another natural or unnatural amino acid and/or protected by one or more substitute organic groups. This is especially true for amino acids L-Orn in position 8.

Thus, in one aspect the present invention provides compounds of formula (1), where in position 8 L-Orn missing.

L-Orn can be replaced by D-Orn, or other natural or neprinol the second amino acid. For example, L-Orn may be substituted natural amino acid such as lysine; or a protected amino acid such as arginine or lysine with one or more alkyl, phenyl or oligometastases substituents, such as N(Me)2N'(Me)2-Arg, N(Me,Ph),N'(Me)2-Arg, N(CH2)4N'(Me)2-Arg, N(CH2)4N'(CH2)4-Arg, Nε(Me)3-Lys.

L-Orn can be protected. For example, the amino group of L-Orn may have substituents, particularly alkyl substituents which may be further substituted, especially heterocyclic groups, for example Nδ(N(CH2)4N'(CH2)4)-Orn; or a more complex substituents, as biotinylation or Orn(NδTfa).

Cyclic amino acids

Other preferred compounds according to this invention include compounds of formula (1), where one or more amino acids of the cyclic chain replaced by other natural or unnatural amino acids protected by organic groups, or removed.

In particular, preferred compounds include compounds with substitution of 1, 2 or 3 amino acids in the cyclic chain.

Especially preferred are such compounds of formula (1), where one or more amino acids are replaced by other natural or unnatural amino acids and/or protected organic groups is. This preferably applies particularly to the amino acids L-Phe at position 3.

Thus, in one aspect the present invention provides compounds of formula (1), where L-Phe at position 3 substituted or protected.

L-Phe may be substituted for D-Phe, or other natural or unnatural amino acid. For example, L-Phe may be substituted natural amino acid such as tyrosine; alkyl substituted by amino, such as N-methyltyrosine; aryl-substituted amino acid, such as 2-amino-3-biphenyl-4-ylpropionic acid, 2-amino-3-naphthalene-2-ylpropionic acid or aminophenylalanine acid; heterocyclizations amino acid, such as 2-amino-3-thiophene-2-ylpropionic acid; or a cyclic amino acid, where the amino group is a heterocyclic part, such as 1,2,3,4-tetraethylene-3-carboxylic acid or octahydrocyclopenta-1-carboxylic acid.

L-Phe can be protected. For example, the phenyl ring may be partially or fully saturated and may be substituted by one or more substituents. The substituents of the phenyl ring can be, as mentioned, preferably, halogen or nitro-group. The amino group may have 1 or 2 substituent, especially alkyl, such as methyl, especially one methyl Deputy.

End acyl group

In addition to modifications of amino acids ecocycles the second chain and/or cyclic portion may also be a connection with a modification in the terminal acyl group ekzoticheskoy chain.

For example, the acyl group may include one or more substituents, especially aromatic, heterocyclic or carbocyclic group, which, in turn, may have one or more substituents, for example, it may be benzoline group which may have one or more substituents, phenylalkanoic, which may have one or more substituents, or cynnamoyl, which may have one or more substituents. In addition, end acyl group can be another essential fatty acid, usually a saturated or unsaturated fatty acid with 3-26 carbon atoms, more preferably 12-20 carbon atoms.

Typical groups for the introduction instead of the terminal acyl group in the formula (1) include:

- linear alkanoyloxy chain with up to 26 carbon atoms and with one or more substituents, especially substituents selected from optionally substituted cycloalkyl, such as 4-methylcyclohexyl; alkyl, especially of short-chain alkyl, such as methyl; optionally substituted aryl, especially phenyl, such as differenl; halogen, such as fluorine up to Perfora; the carbonyl group, amino group or aminogroup;

- optionally substituted benzoyl, such as the benzoyl, p-methylbenzoyl, p-trifloromethyl, piperonal;

- arylalkylamine group, preferably is, with the two carbon atoms in alkanoyl, especially cinnamoyloxy group, such as p-cryptomaterial.

In addition, the acyl group may be replaced by another acyl group, preferably a group of the formula R CO-. R' means the above-described radical and means, respectively, alkyl, alkenyl, aryl, heterocyclyl or carbocyclic itself can be replaced.

Finally, it can be connect with the only modification being in fatty acid ekzoticheskoy circuits for the formation of 5-methylhexane end of the group.

Combined replacement

Possible replacement in ekzoticheskih amino acids, cyclic amino acids and the terminal acyl group can be carried out in combination.

Thus, other preferred compounds according to this invention are such compounds of formula (1), where one or more amino acids ekzoticheskoy chain and/or one or more amino acids of the cyclic part is modified, as described above, and/or modified terminal acyl group.

Compounds where L-Orn at position 8 is replaced or protected, may have other preferred modifications, including 4(S)-methylhexane or another group in the terminal group of the side chain.

Compounds where L-Phe at position 3 substituted or protected, may have other prefer the performance communications modifications including 4(S)-methylhexane or another group in the terminal group of the side chain.

Examples of substituting groups include1-C18-alkyl, C2-C18alkenyl,2-C18-quinil, aryl, heterocyclic group, OR', SR', SOR', SO2R', NO2That other', N(R')2, NHCOR', N(COR')2, NHSO2R', CN, halogen, C(=O)R', CO2R', OC(=O)R'where each R'group is independently selected from the group consisting of H, HE, NO2, NH2, SH, CN, halogen, C(=O)H, C(=O)-alkyl, CO2H, substituted or unsubstituted With1-C18-alkyl, substituted or unsubstituted With2-C18-alkenyl, substituted or unsubstituted With2-C18-quinil and substituted or unsubstituted aryl. Suitable halogen-substituents in the compounds according to the present invention include F, Cl, Br and I.

Alkyl groups are saturated linear or branched hydrocarbon group including, for example, methyl, ethyl, isopropyl, isobutyl, tert-butyl, heptyl, dodecyl, octadecyl, amyl, 2-ethylhexyl, 2-methylbutyl, 5-methylhexan, 9-methyl-3-decyl and the like, in particular alkyl groups which have one side methyl group. Suitable alkyl group is long and has 1-20 carbon atoms, more typically 1-15 or 1-10 carbon atoms, or may be short and has 1-6 or 1-3 atom of carbon is and. Long carbon chain are "candidates" for use in the terminal group of the fatty acid.

Preferred alkeneamine and alkyline group in the compounds according to the present invention have one or more unsaturated linkages and from 2 to 12 carbon atoms, more preferably 2-8 carbon atoms, more preferably 2-6 carbon atoms, even more preferably 2, 3 or 4 carbon atoms. The terms "alkenyl" and "quinil", as used in this context include both cyclic and acyclic groups, although linear or branched acyclic groups are usually preferred. In a General sense, the authors include the term "alkylidene" in the term "alkenyl", because they both represent substituents with a double bond.

Suitable aryl groups in the compounds according to the present invention include monocyclic or polycyclic compounds, including polycyclic compounds that contain separate and/or condensed aryl group. Typical aryl groups contain 1 to 3 separate or condensed cycles and 6-18 carbon atoms in cycles. Particularly preferred aryl groups include substituted or unsubstituted phenyl, naphthyl, biphenyl, tenantry and antracol.

Suitable acyl groups include alcoholnye groups that have -12 carbon atoms, more preferably 2-8 carbon atoms, more preferably 2-6 carbon atoms, even more preferably 2 carbon atoms. Other acyl groups include alkenylacyl, alkenylacyl, arylaryl, heterocyclyl.

Suitable heterocyclic groups include heteroaromatic and heteroalicyclic group. Suitable heteroaromatic groups in the compounds according to the present invention contain one, two or three heteroatoms selected from the atoms N, O or S, and include, for example, coumarinyl, including 8-coumarinyl, chinoline, including 8-chinoline, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl, benzofuranyl and benzothiazole. Suitable heteroalicyclic group in the compounds according to the present invention contain one, two or three heteroatoms selected from the atoms N, O or S, and include, for example, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholinopropan and pyrrolidinyloxy group.

The above groups may be substituted at one or more available positions by one or more substituents.

Natural amino acids include alanine, glycine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, methionine, cysteine, aspartate, asparagine, glutamic acid, glutamine, lysine, arginine, Proline, serine, threonine, histidine is hydroxyproline.

The authors also refer to WO 0158934 fully included in this context as a reference. The above text is a guide that is applicable to compounds according to this invention, and the authors introduce the definitions used in the case of compounds according to this invention.

Abbreviations used in the following description for amino acids and peptides follow the rules of the Commission on biochemical nomenclature of the IUPAC-IUB according to J. Biol. Chem., 1972, 247, 977-983.

Use the following additional abbreviations:

Allocallyloxycarbonyl
N(CH2)4N'(CH2)4-Arg2-amino-5-[(dipyrrole-1-ylmethylene)amino]pentane acid
AMminiate acid
JSColaoluwa acid
Bip2-amino-3-biphenyl-4-ylpropionic acid
Vostert-butyloxycarbonyl
BTFFHbis(tetramethylene)performative-hexaphosphate
t-Butert-butyl
Bza-OHbenzoic acid
Chacyclohexylamin or 2-amino-3-cyclohexylpropionic acid
p-CF3Bza-OH4-triftorperasin acid
p-CF3Bz-OH3-(4-trifloromethyl)acetic acid
p-CF3Spp-OH3-(4-trifloromethyl)acrylic acid
Cl-TrtCl-resin2-horticulturally resin
Dha2-aminoacridone acid or didigitally
Z-Dhbthe α,β-didehydro-α-aminobutyric acid
DIPEAN,N-diisopropylethylamine
DMFN,N-dimethylformamide
EDC·HCl1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride
Fmoc9-fluorenylmethoxycarbonyl
6,6-dFHep-OH6,6-differetnly acid
3,5-dFPhAc-OH(3,5-differenl)acetic acid
4-GuBut-OH4-guanidinopentanoic acid
GI50inhibition of growth by 50%
HAPyUO-(7-asobancaria-1-yl)-1,1,3,3-bis(tetramethylene)orangestriped
HATUO-(7-asobancaria-1-yl)-1,1,3,3-tetramethyleneglutaric
hChhomecollection or 2-amino-4-cyclohexylaniline acid
Hep-OHheptane acid
SPLAacetic acid
HOAt1-hydroxy-7-asobancaria(3-hydroxy-3H-1,2,3-triazolo[4,5-b]pyridine)
HOBt1-hydroxybenzotriazole
Icos-OHCasanova acid
LC50lethal to 50% concentration
Lit-OHlithocholic acid
Maneh-HEmethylhexanoic acid
M2And[(tetramethylene)-1H-1,2,3-triazolo[4,5-b]pyridine-1-ylmethylene]-N-methylmethanesulfonate
(with a/t-4Me-cHexa)-OH(CIS/TRANS)-4-methylcyclohexanecarboxylic acid
p-MeBza-OH4-methylbenzoic acid
N(Me)2N'(Me)2-Arg2-amino-5-(N',N',N”,N”-tetramethylguanidine)pentane acid
N(CH2)4N'(Me)2-Arg2-amino-5-[(dimethylaminopyridine-1-ylmethylene)amino]pentane acid
Meonmethanol
N(Me,Ph),N'(Me)2-Arg2-amino-5-(N',N',N'-trimethyl-N-phenylquinoline)pentane acid
NMMN-methylmorpholin
Mst-OHmyristic acid or tetradecanoic acid
NaI2-amino-3-nafta is in 2-ylpropionic acid
Oct-OHoctanoic acid
6-Oner-HE6-oxyptera acid
Oicoctahydrocyclopenta-1-carboxylic acid
Nδ(N-(CH2)4-N(CH2)4)-Orn2-amino-5-[(dipyrrole-1-ylmethyl)amino]pentane acid
Phf-OHperformative acid
Phgphenylglycine or aminophenylalanine acid
(5R)-Ph-Pro5-(R)-phenylpyrrolidine-2-carboxylic acid
Pippipecolinate acid
Rre-HEbenzo[1,3]dioxol-5-carboxylic acid
SPSsolid phase synthesis
TfaTRIFLUOROACETYL
TFAtriperoxonane acid
TFAtriperoxonane anhydride
TGIcomplete growth inhibition
ThiAla-[3-(2-thienyl)] or 2-amino-3-thiophene-2-ylpropionic acid
Tic1,2,3,4-tetraethylene-3-carboxylic acid
Und-OHundecanoate acid

Denote the amino acids specified in the L-configuration except where indicated otherwise.

Examples of compounds according to this invention include compounds of formula (1)where:

Is Glu or Lys is instead of L-Orn at position 8;

- Gly, Phe, Ala, Leu, D-Val, Pro, Gln, Orn, Thr, or Glu are instead of L-Val at position 11;

- Val or D-Thr is instead of L-Thr at position 12;

- Gly, D-Ala, D-Leu, D-Phe, D-Pro, Val, D-Glu, D-Gln or D-Thr are instead of D-Val at position 13;

- hCh is instead of L-Val at position 11 and/or D-Cha is instead of D-Val at position 13;

- Ala, Gly, Leu, Pro, Glu, Orn or Gln are instead of L-Thr at position 12 and no amino acid at position 13;

- D-Ile or D-Val is instead of D-allo-Ile at position 7;

- D-Orn is instead of L-Orn at position 8 and L-Pro instead of D-Pro at position 9;

- D-Cys is instead of D-allo-Ile at position 7 and L-Cys is instead of L-Val at position 11; or

- D-Cys or D-homo-Cys is instead of D-allo-Ile at position 7 and D-Cys or D-homo-Cys is instead of D-Val in position 10.

- Icos, (ct)-4-Me-cHexa, Und, (4R)-MeHex, (4RS)-MeHex, (4S)-MeHex, Oct, p-MeBza, Bza, p-CF3Bza, 3,5-dFPhAc, Pipe, p-CF3Cinn, p-CF3PhAc, Pfh, 6-OHep, 6,6-dFHep, 4-GuBut, AM, AO, or C(=N(CH3)2are instead of 5-MeHex at position 14 and, optionally, Lys is instead of L-Orn at position 8;

- Pro, D-Pip, D-Tic, or (5R)-Ph-Pro are instead of D-Pro at position 9 and (4S)-MeHex is instead of 5-MeHex in position 14;

- N(Me)2N'(Me)2-Arg, N(Me,Ph),N'(Me)2-Arg, N(CH2)4N'(Me)2-Arg, N(CH2)4N'(CH2)4-Arg, Nδ(N(CH2)4N'(CH2)4)-Orn, Nε(Me)3Is Lys, Orn(NδTfa) or Orn(Biot) are instead of L-Orn at position 8, and, optionally, (4S)-MeHex is instead of 5-MeHex at position 14, and, optionally, Thr(OTfa) is instead of L-Thr at position 12;

- Thr(OTfa) is instead of L-Thr at position 12 and (4S)-MeHex or Lit(OTfa) is instead of 5-MeHex in position 14;

- N-(Hep)2-D-Val is instead of D-Val at position 13 and there is no 5-MeHex in position 14; or

- there are no amino acids at positions 11, 12 and 13 and, optionally, Mst is instead of 5-MeHex in position 14.

- Dha or E-Dhb is instead of Z-Dhb at position 2;

- D, L-Ser, Gly or Aib are instead Thr at position 2, and are hydrogenated analogs;

- D-Val is instead of L-Val at position 1;

- Trp is instead of L-Phe at position 3; or

- D-Thr or D-Ser is instead of D-alo-Thr at position 6.

- hCh, Phe(3,4-Cl2), Phe(F5), Phe(4-I), Phe(4-NO2), Phe(4-F), Tyr(Me), Thi, Tic, Tyr, Oic, NMePe, Phe(2-Cl), Phe(3-Cl), Phe(4-Cl), Phe(3,4-F2), NaI, Bip or Phg are instead of L-Phe in position 3 and, optionally, (4S)-MeHex or p-CF3Cinn is instead of 5-MeHex in position 14;

- D-Val or D-Cha is instead of D-alo-Ile at positions 5 and 7 and, optionally, D-Cha is instead of D-Val at position 4; or

- D-Val is instead of L-Val at position 1, D-Phe is instead of L-Phe at position 3, Val is instead of D-Val at position 4, L-alo-Ile is instead of D-alo-Ile at position 5, L-alo-Thr is instead of D-alo-Thr at position 6, L-alo-Ile is instead of D-alo-Ile at position 7, D-Orn is instead of L-Orn at position 8, L-Pro instead D-Pro at position 9, L-Val is instead of D-Val at position 10, D-Val is instead of L-Val at position 11, D-Thr is instead of L-Thr at position 12 and L-Val is instead of D-Val in position 13.

Accordingly, these proposed replacement can be performed in combination.

The present invention also covers their pharmaceutically acceptable salts, prodrugs, tautomers and solvate.

Compounds according to the present invention have asymmetric centers and therefore exist in different enantiomeric and diastereomeric forms. This invention relates to the use of all optical isomers and stereoisomers of the compounds according to the present invention and their mixtures, and to all pharmaceutical compositions and methods of treatment that can be applied or which can is activated.

Compounds according to this invention may be in crystalline form or in the form of the free compounds or in the form of a solvate (e.g. hydrate) and assume that both forms are within the scope of the present invention. Methods of solvation, generally known in the prior art.

The present invention also includes compounds according to the present invention and their pharmaceutically acceptable salts, where one or more atoms of hydrogen, carbon or other atoms are replaced by their isotopes. Such compounds may be useful as research and diagnostic tools in pharmacokinetic study of metabolism and analyses on binding.

As stated in the description, the compounds according to this invention, including the compounds of formula (1)include their pharmaceutically acceptable derivatives or prodrugs. The term "pharmaceutically acceptable derivative or prodrug" means any pharmaceutically acceptable salt, ester, salt of ester or other derivative compounds according to this invention that when administered to a recipient is able to give (directly or indirectly) the compound according to this invention, or a metabolite or residue. Especially preferred derivatives and prodrugs are those that increase b is dostupnost compounds according to this invention, when such compounds are administered to a patient (for example, given that the oral input connection easier absorbed in the blood), increase the delivery of the parent compound in a given biological compartment, increase solubility for administration by injection, alter metabolism or alter the rate of excretion.

Salts of the compounds according to the present invention may include an additive salts of the acids formed by the nitrogen atom in the compound of formula (1) or (2). Therapeutic activity inherent part deriving from the compounds according to this invention, as specified here, and the identity of the other component is of less importance although for therapeutic and prophylactic purposes it is preferably pharmaceutically acceptable for the patient. Examples of pharmaceutically acceptable additive salts of acids include those derived from mineral acids such as hydrochloric acid, bromatologia acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid; and organic acids such as tartaric acid, acetic acid, triperoxonane acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid, methanesulfonate acid and Arinsal is about acid, for example, p-toluensulfonate acid. Preferred salts include triptorelin and hydrochloride.

Compounds according to the present invention can be obtained according to the synthesis method described in WO 0158934, or in accordance with the improved method, as described in the description. Therefore, this invention also relates to a method for obtaining compounds corresponding to the formula (1).

Key steps in the optimized way for more efficient and safe synthesis kahalalide F and its analogues are: (i) partial inclusion of Fmoc-D-Val-OH in horticulturally resin with an initial loading of 0.5 mmol/g; (ii) use as a way of linking DIPCDI-HOBt instead of HATU-DIPEA to sequentially activate the protected amino acids and aliphatic carboxylic acids; (iii) stage cyclization DIPCDI/HOBt/DIPEA in CH2Cl2; in these conditions, avoid two adverse reactions: the epimerization of the Val residue, which is involved in the activation and triptoreline Phe or its replacement; (iv) the use of sodium diethyldithiocarbamate after removal of the Alloc, to avoid the presence of Pd(O) in the final product.

The synthesis is, preferably, solid-phase synthesis.

The preferred embodiment of the method of synthesis according to the present invention is best shown in figure 1, which relates the I to the formation of the target compounds.

Scheme 1

As shown above in scheme 1, the preferred synthetic method of education analogues kahalalide F based on solid-phase method, see, for example, Lloyd-Williams P., and others, Chemical Approaches to the Synthesis of Peptides and Proteins. CRC Press, Boca Raton (FL), 1997, and following the modifications of the method already described for obtaining kahalalide F and some of its analogues (WO 0158934).

The method in scheme 1 comprises the following stages:

(a) the introduction of Fmoc-D-Val-OH in horticultural with the formation of ester bonds;

(b) the elongation of the peptide chain from three amino acids [concert, DaThr(free OH), concert] with the use of strategy Fmoc/tBu;

(C) the introduction of [Val(1)] with the use of strategy Alloc/tBu;

(d) elongation of the peptide chain due to residual amino acids and aliphatic carboxylic acids with the use of strategy Fmoc/tBu;

(E1) the introduction of the dipeptide Alloc-Phe-ZDhb-OH, which combine and dehydration in solution;

(EA) elongation of the peptide chain by two amino acids, preferably, Thr and Phe. The group HE amino Thr is unprotected, and the amino group of Phe, or its replacement, protected with Fmoc or, preferably, using Alloc; in some cases, if it is protected with Fmoc group, it is removed and Alloc is introduced into the solid phase;

(2b) dehydration in the solid phase to obtain delegitimated;

(f) removal of Alloc/Fmoc-Phe group, or it is ameny, while the peptide is still attached to the solid substrate;

(g) removal is protected in the side chain of the peptide from the solid foundations;

(h) the cyclization of the peptide in solution;

(i) removing TFA labile protective groups of the side chain;

(j) further modification of the functional(Oh) group(s) in the solution phase.

Therefore, the method can be performed as follows:

Fmoc-D-Val-OH is injected, preferably in horticulturally resin, see Barlos, K.; Gatos, D.; Schäfer, W., Angew. Chem. Int. Ed. Engl., 1991, 30, 590-593, in the presence of DIPEA, while maintaining a degree of substitution of about 0.5 mmol/g, the Application of higher loads cause the presence of terminal peptides in the final product, see Chiva C.; Vilaseca, M.; Giralt, E.; Albericio F.; J. Pept. Sci., 1999, 5, 131-140.

All the relative costs of solvents are given.about. unless specified otherwise.

Removal of Fmoc groups are carried out with piperidine-DMF (2:8, vol/about.)(1×2 min, 2×10 min). Binding Fmoc-AA-HE (4-5 equiv.) and acid at position 14 can be accomplished using DIPCDI-HOBt (equimolar amount of each of them belong to the carboxyl component) or PyBOP-DIPEA (equimolar amount of PyBOP and double the number of DIPEA) in DMF or DMF-toluene (1:1) within 90 minutes After bonding exercise ninhydrin or chloranilic tests and, if the test is positive, repeat binding under the same conditions, otherwise, %the SS continue. Washing between stages of the removal of protection, binding, and again remove protection are carried out with DMF (5×0.5 min) and CH2Cl2(5×0.5 min), using, for example, each time with 10 ml of solvent per gram of resin.

Introduction Alloc-Val-OH (5 EQ.) can be carried out with equimolar amounts of DIPCDI and 10% DMAP. This link is repeated at least twice.

The removal of the Alloc group can be accomplished using Pd(PPh3)4(0.1 EQ.) in the presence of PhSiH3(10 equiv.) see Gómez-Martínez, P.; Thieriet N.; Albericio F.; Guibé F.; J. Chem. Soc. Perkin I, 1999, 2871-2874, and washing the resin 0.02 M solution of sodium diethyldithiocarbamate in DMF (3×15 min).

Dipeptide Alloc-Phe-ZDhb-OH (4 equiv.) which is obtained in the solution of Alloc-Phe-HE and H-Thr-OtBu with EDC·HCl, followed by dehydration and processing of TFA can be associated with DIPCDI-HOt (4 equiv. each) for 5 hours to overnight. The use of other reagents binding, based on HOt, such as HBTU or DIPCDI-HOt, leads to incomplete inclusions of the dipeptide.

Dehydration can be accomplished in the solid phase using EDC·HCl (water-soluble carbodiimide, 20 EQ.) in the presence of CuCl (12 EQ.) in CH2Cl2-DMF (9:1) within 7 days. EDC·HCl/CuCl used for dehydration in the solution of the Thr residue in the fragment of lowland (Fukase, K.; Kitazawa, M.; Sano, A.; Shimbo, K.; Horimoto, S.; Fujita, H.; Kubo, A.; Wakamiya, T.; Shibe A. Bull. Chem. Soc. Jpn., 1992, 65, 2227-2240) and in the solid phase to obtain delegitimation from Thr, Se and phenylsilane (Royo, M.; J.C. Jiménez; López-Maciá, A.; Giralt, E.; Albericio F.; Eur. J. Org. Chem., 2001, 45-48).

Cleavage of the protected peptide from the resin can be accomplished using TFA-CH2Cl2(1:99)(5×30 sec).

Stage cyclization can be accomplished using DIPCDI/HOBt/DIPEA in CH2Cl2. Under these conditions, avoid two adverse reactions: the epimerization of the Val residue, which is involved in the activation and triptoreline Phe or his replacement.

The final destruction of the protection can be accomplished using TFA-H2Oh (95:5) for 1 hour.

You should take into account that the specific choice of protective groups is not critical and widely available alternatives. For example, groups such as Bzl, you can replace tBu/Boc; Boc instead of Fmoc; Fmoc instead of Alloc; Wang resin instead chlorotrityl.

Further details of the synthesis are given in the examples.

The method according to this invention can be made from raw materials of enantio-, stereocontrolled and quickly done by using the advantages of solid-phase synthesis methodology, where the molecule in the design associated with an insoluble basis for all operations of synthesis.

Pharmaceutical finished dosage forms of the compounds according to this invention can be adapted for administration by any appropriate route, such as oral (including buccal or sublingual), rectal, natalin is m, local (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) route. Such medicines can be obtained by any method known in the field of pharmacy, for example by Association the active ingredient with the carrier(s) or excipient(s).

Preferably, pharmaceutical compositions containing the compounds according to this invention include liquid compositions (solutions, suspensions or emulsions) with suitable composition for intravenous, and they may contain the pure compound or be in combination with any carrier or other pharmacologically active compounds. Further guidance regarding the pharmaceutical compositions can be found in WO 0236145, which is fully included in this context by reference.

Thus, the combination of nonionic surfactants and organic acids suitable for use with a filler to obtain a freeze-dried forms of the compounds according to this invention, suitable for recovery. Recovery is preferably carried out in a mixture of emulsifying a solubilizer, alkanol and water.

Freeze-dried composition preferably includes, mainly, fill the l, for example, at least 90% or at least 95% filler. Examples of fillers are well known and include sucrose and mannitol. You can use other fillers.

Nonionic surface-active substance in the dried composition is, preferably, a complex arbitarily ether, more preferably, polietilensorbit ester, such as polyoxyethylenesorbitan, especially a polyoxyethylene sorbitan monooleate such as Polysorbate 80. Nonionic surfactant is typically a few percent of the composition, as, for example, 0-5% of the composition, for example 2, 3 or 4% of the composition.

Organic acid in the dried composition is typically an aliphatic acid, preferably hydroxycarbonate acid and, more preferably, hydroxypolycarboxylic acid, especially citric acid. Organic acid is typically a few percent of the composition, as, for example, 0-5% of the composition, for example 2, 3 or 4% of the composition.

The number of compounds according to this invention in a freeze-dried composition is usually less than 1% or, more commonly, less than 0.1%, based on the mixture. A suitable amount is in the range of 50-200 μg, about 100 μg to 100 mg of the composition.

Emulsifying the solubilizer for stanowiacego agent, accordingly, includes an ester of polyethylene glycol, especially the fatty acid ester, more preferably, PEG oleate, such as oleate PEG-35. Emulsifying the solubilizer is, accordingly, 0-10% recovery agent, usually about 3-7%, approximately 5%. Alkanol usually is ethanol and is, accordingly, 0-10% recovery agent, usually about 3-7%, approximately 5%. The rest of the recovery agent is water, which receive recovery solution suitable for intravenous injection.

Further dilution recovery solution with 0.9%saline solution may be appropriate for infusion kahalalide connection. Suitable equipment for infusion preferably includes a glass container, which is more preferable than that of polyethylene. The tube preferably is silicone.

The preferred reduction agent then contains 2-7%, approximately 5%, emulsifying a solubilizer; 2-7%, approximately 5%, of alcohol; and the rest is water.

Finished dosage forms can be represented in the form of packages with standard dose or multiple doses, for example sealed ampoules and vials and may be stored in subjected to drying by freezing (lipelis is proofed), immediately prior to use, requiring only the addition of sterile liquid carrier, for example water for injection.

This invention additionally provides a pharmaceutical kit containing a separate container filled with freeze-dried composition and reducing agent. Also provides methods of recovery.

Introduction compounds or compositions according to the present invention is carried out by intravenous infusion. Time of infusion could be up to 72 hours, more preferably up to 1-24 hours, or most preferably up to about 1 or up to about 3 hours. Especially desirable short-term infusion, which give the opportunity to carry out treatment all night outside the clinic. However, the infusion can be done about 24 hours or even longer if necessary.

Introduction perform the cycles used by the preferred method; intravenous infusion of the compounds according to this invention, patients are subjected in the first week of each cycle and the patients are able to recover during the remainder of the cycle. The preferred duration of each cycle is 1 or 3 or 4 weeks; multiple cycles can be performed as needed. In alternative dosing Protocol connection according to this izobreteny is introduced in the space of about 1 hour for 5 consecutive days for 3 weeks. Other protocols can be as variations.

Slow dispensing and/or reducing the dosing and adjustment circuits applications perform as required, depending on the individual tolerance of the patient treatment, in particular, reduction of dosage is recommended for patients with higher than normal levels of serum hepatic transaminases or alkaline phosphatase.

In one aspect the present invention relates to a method for the treatment of human cancer, including the introduction of the above to a patient compounds according to this invention in a dose of less than 1200 micrograms twice a day, preferably less than 930 µg twice a day and, more preferably, less than 800 mcg twice a day. Suitably, the dose is at least 320 mcg twice a day. Preferably, the dose is in the range of 400-900 mcg twice daily, preferably 500-800 mcg twice daily, more preferably 600 to 750 mcg twice a day. Especially preferred is a dose of approximately 650-700 mcg twice a day.

In a further aspect this invention relates to a method for the treatment of human cancer, including the introduction of the above patient connection according to this invention, daily for 5 days at a dose of less than 930 µg twice daily, with the last is the missing break from 1 to 4 weeks, in which kahalalide compound is not administered. The dose is preferably 650-750 mcg twice daily, more preferably, about 700 μg, twice a day. The time of infusion is preferably 1-24 hours, more preferably 1-3 hours. Particularly preferably, the time of infusion is about 1 or about 3 hours. Break is preferably 2-3 weeks, more preferably, about 2 weeks.

The present invention also relates to a method for treatment of human cancer, including the introduction of the above patient connection according to this invention once a week in doses of less than 800 mcg twice a day. The dose is preferably 600-700 μg twice daily, more preferably, 650 mcg twice a day. The time of infusion is preferably 1-24 hours, more preferably 1-3 hours.

Although the instructions on the dosage given above, the exact dosage of the compounds will vary according to the particular finished dosage form, method of application and the specific location of the recipient and the treated tumor. Other factors to consider are age, body weight, sex, diet, time of administration, rate of excretion, condition of the recipient, combination of drugs, sensitivity to the effects of the severity of the disease. The introduction is carried out continuously or intermittently, within the maximum permissible dose.

The present invention particularly relates to the treatment of patients affected by prostate cancer, breast cancer, hepatocellular cancer, melanoma, cancer of the colon and rectum, kidney cancer, ovarian cancer, NSCL cancer, epithelial cancer, pancreatic cancer and tumors, which sverkhekspressiya the oncogene Her2/neu. More preferably, it relates to the treatment of hepatocellular cancer, melanoma, breast cancer, pancreatic cancer and prostate cancer.

The present invention also relates to a method for treatment of skin diseases caused by hyperproliferative skin cells in a mammal which comprises the administration to a mammal an effective non-toxic amount of the compound according to this invention. Skin disease, preferably, is psoriasis. The present invention relates preferably to the treatment of people affected with psoriasis, particularly severe form of psoriasis.

The compounds and compositions according to this invention can be used with other medicines to combined therapy. Other medicines can also be part of the same composition, or be provided as a separate composition and to introduce at the same time or at another time. The identity of another drug is not particularly limited, although it is envisaged combination with other chemotherapeutic agents, hormonal agents or antibodies. The number of compounds according to this invention and the other pharmaceutically active agent or agents and the relative regulation of the target in order to achieve the desired combined therapeutic effect.

EXAMPLES

Common procedures

Cl-TrtCl-resin-protected Fmoc-amino acid derivatives, HOBt, HOAt are issued by firms ABI (Framingham, MA), Bachem (Bubendorf, Switzerland), NovaBiochem (Läufelfingen, Switzerland), 4-Menej-derivatives - the company Narchem; HATU and other handlerbase derivatives are produced by the firm ABI (Framingham, MA), or get them according to M. del Fresno, El-Faham, A.; L.A. Carpino; Royo, M.; Albericio F. Organic Lett., 2000, 2, 3539-3542.

Alloc-amino acids, in essence, receive, as described Dangles and others, see Dangles O.; Guibé F.; Balavoine, G.; Lavielle, S.; Marquet. A. J. Org. Chem., 1987, 52, 4984-4993; and Alloc-Z-Dhb-Phe-OH and kahalalide F receive, as described in WO 0158934; DIPEA, DIPCDI, EDC·HCl, piperidine, TFA issued by the company Aldrich (Milwaukee, WI). DMF and CH2Cl2produced by the firm SDS (Peypin, France). Acetonitrile (purity grade for HPLC) manufactured by a company Scharlau (Barcelona, Spain). All commercial reagents and solvents used as received, except for CH2Cl2that p is lowered through a column of alumina to remove impurities of acid character.

Solid phase synthesis was carried out in polypropylene syringes (10-50 ml), equipped with a porous polyethylene disk. Solvents and soluble reagents are removed by suction. Removal of Fmoc groups are carried out with piperidine-DMF (2:8, vol/about.) (1×2 min, 2×10 min). Washing between stages of the removal of protection, binding, and again remove protection are carried out with DMF (5×0.5 min) and CH2Cl2(5×0.5 min), using each time 10 ml of solvent per gram of resin. Turning peptide synthesis and washing performed at 25aboutC. Syntheses carried out on solid phase, monitored by HPLC intermediate products obtained after cleavage with TFA-H2O (1:99) for 1 min in the aliquot (approximately 2 mg) peptidyl resin. HPLC column reversed-phase [Nucleosil-C18a 4.6×250 mm, 10 μm (column A); Nucleosil-C4a 4.6×250 mm, 10 μm (column C) are issued by the firm Sharlau, Spain); Symmetry™ C18a 4.6×150 mm, 5 μm (column C); Symmetry300™ C18a 4.6×50 mm, 5 μm (column D) are issued by the firm Waters (Ireland), and Bond SB C18a 4.6×150 mm, 3.5 µm (column E) issued by the company Agilent (USA)]. Analytical HPLC is carried out on a Waters instrument containing two pressure pump for solvent (Waters 1525), an automatic injector (auto sampler, Waters 717), the detector for the two wavelengths (Waters 2487) and system controller (Breeze V3.20; and on the instrument Agilent 1100 containing two pressure pump for solvent (G1311), an automatic injector (G1329), DAD (G1315). UV detection was performed at 215 or 220 nm, and linear gradients CH3(CN+being 0.036% TFA) in N2About (+0,045% TFA) operate in the following conditions:

Table I
Chromatographic conditionsFlow (ml/min)Gradient
(% CH3CN)
Duration (min)
A1,030-10030
In1,040-6015
1,045-608
D1,040-708
E1,045-708
F1,040-7015
G1,040-6515
H1,010-10030
I1,045-6515
J1,040-7015
To1,055-7515
L1,040-10015
M1,050-1008
N1,035-6015
About1,050-10030
P1,050-10015
Q1,0sacratissimi 45 15
R1,030-10015
S1,020-1008
T0,635-9025
U0,740-7050
V0,810-48 for 15 min and isocratic 30 min45
W1,010-7050
X1,010-48 for 15 min and isocratic 30 min45
Y0,810-6050
Z0,715-7055
AB0,810-60 55
AC0,830-6045
AD0,810-48 for 10 min and isocratic 30 min40
AE1,045-9025
AF1,040-1008

Analysis by time-of-flight mass spectrometry with laser desorption and ionization of the matrix (MALDI-TOF) mass spectrometry with ionization electron spray (ES-MS) peptide samples carried out on a PerSeptive Biosystems Voyager DE RP, using DHB matrix, and spectrometer Waters Micromass ZQ, and Agilent Ion Trap 1100 series LC/MSDTrap. Samples containing the peptide resin hydrolyzing in a mixture of 12 N. aqueous HCl-propionic acid (1:1) at 155aboutC for 1-3 hours and not containing peptide samples hydrolyzing 6 N. aqueous HCl at 155aboutC for 1 hour. Subsequent amino acid analysis performed on the auto-analyzer Beckman System 6300.1H-NMR-spectroscopy [1H, NOESY, TOCSY if (C)] carried out on a Varian Unity Plus 500 MHz). Chemical shifts (δ), measured from the S towards less strong field, expressed the indicate in ppm (ppm). Constant interaction is expressed in Hertz.

Denote the analogues do about 5-methylhexane isomer kahalalide F formula (I), indicating in brackets the modified residue; the suffix "no" refers to the exclusion of the natural balance of the sequence.

Example 1

(4S)-MeHex-D-Val-ThrVal-D-Val-D-Pro-Orn-D-allo-Ile-cyclo[D-allo-Thr-D-allo-Ile-D-Val-Phe-(Z)Dhb-Val], [(4S)-MeHex14]-kahalalide F(compound 1)

Stage 1

H-D-Val-O-TrtCl-resin

Cl-TrtCl-resin (1 g, of 1.64 mmol/g) was placed in a 20 ml polypropylene syringe fitted with a polyethylene filter disk. The resin is then washed with CH2Cl2(5×0.5 min) and add a solution of Fmoc-D-Val-OH (238 mg, 0.7 mmol, 0.7 EQ.) and DIPEA (0,41 ml) in CH2Cl2(2.5 ml) and the mixture stirred for 15 min, then introduce additional DIPEA (0,81 ml, only 7 EQ., 7 mmol) and the mixture is stirred for 45 minutes Interaction complete addition of the Meon (800 ml) after stirring for 10 minutes Fmoc-D-Val-O-TrtCl-resin is subjected to subsequent leaching/treatments CH2Cl2(3×0.5 min), DMF (3×0.5 min), piperidine, as described in the General methods, and DMF (5×0.5 min). Loading calculated by determining the Fmoc, is 0.50 mmol/g

Stage 2

Fmoc-D-allo-Ile-D-allo-Thr(Val-Alloc)-D-allo-Ile-D-Val-O-TrtCl-resin

Fmoc-D-allo-Ile-HE (707 mg, 2 mmol, 4 equiv.) Fmoc-D-allo- Thr-HE free the Naya hydroxyl group) (683 mg, 2 mmol, 4 equiv.) and Fmoc-D-allo-Ile-HE (707 mg, 2 mmol, 4 equiv.) consistently add to viseporodicnog H-D-Val-O-TrtCl-resin using DIPCDI (310 μl, 2 mmol, 4 equiv.) and HOBt (307 mg, 2 mmol, 4 equiv.) in DMF (2.5 ml). In all cases, after 90 min the reaction of binding of the ninhydrin test is negative. Removal of Fmoc group and rinsing is carried out, as described in the General methods. Alloc-Val-OH (502 mg, 2.5 mmol, 5 equiv.) associated with DIPCDI (387 mg, 2.5 mmol, 5 equiv.) in the presence of DMAP (30,6 mg, 0.25 mmol, 0.5 EQ.) and DIPEA (88 μl, 0.5 mmol, 1 EQ.) within 45 minutes of This binding is repeated in the same conditions twice. An aliquot peptidyl resin is treated with TFA and HPLC (tR7,8 min, condition S, column D) of the crude product obtained after evaporation shows a purity >98%.

ES-MS calculated for C45H63N5O11: 849,45.

Found: m/z 850,1 [M+H]+.

Stage 3

Fmoc-D-Val-D-Pro-Orn(Boc)-D-allo-Ile-D-allo-Thr(Val-Alloc)-D-allo-Ile-D-Val-O-TrtCl-resin

Fmoc group is removed and Fmoc-Orn(Boc)-HE (912 mg, 2 mmol, 4 equiv.) Fmoc-D-Pro-HE (843 mg, 2.5 mmol, 5 equiv.) and Fmoc-D-Val-HE (255 mg, 2.5 mmol, 5 equiv.) successively added to the above peptidyl resin (stage 2)using DIPCDI (310 μl, 2.0 mmol) and 4 equiv.; and 388 μl, 2.5 mmol, 5 equiv.) and HOBt (307 mg, 2.0 mmol) and 4 equiv.; and 395 mg, 2.5 mmol, 5 equiv.) within 90 minutes Ninhydrin test after the introduction of the Orn and D-Pro is negative. Chloranilic test after the introduction the of D-Val is slightly positive, and, therefore, re-linking this residue with an Fmoc-D-Val-HE (678 mg, 2.0 mmol, 4 equiv.) DIPCDI (310 μl, 2.0 mmol, 4 equiv.) and HOBt (307 mg, 2.0 mmol, 4 equiv.) within 90 minutes an Aliquot peptidyl resin is treated with TFA and HPLC (tR=10,1 min, condition S, column D) of the crude product obtained after evaporation shows a purity >98%.

MALDI-TOF-MS calculated for C65H97N9O16: 1259,71.

Found: m/z 1282,16 [M+Na]+.

Stage 4

(4S)-MeHex-D-Val-Thr(tBu)-Val-D-Val-D-Pro-Orn(Boc)-D-allo-Ile-D-allo-Thr(Val-Alloc)-D-allo-Ile-D-Val-O-TrtCl-resin

Fmoc group is removed and Fmoc-Val-HE (678 mg, 2 mmol, 4 equiv.) Fmoc-Thr(tBu)-HE (992 mg, 2.5 mmol, 5 equiv.) Fmoc-D-Val-HE (678 mg, 2 mmol, 4 equiv.) and (4S)-MeHex-HE (195 mg, 1.5 mmol, 3 EQ.) successively added to the above peptidyl resin (stage 3), using DIPCDI (233 μl, 1.5 mmol and 3 EQ.; and 310 μl, 2 mmol) and 4 equiv.; and 388 μl, 2.5 mmol, 5 equiv.) and HOBt (230 mg, 1.5 mmol and 3 EQ.; 307 mg, 2 mmol) and 4 equiv.; and 395 mg, 2.5 mmol, 5 equiv.) within 90 minutes In all cases after 90 min of binding ninhydrin test is negative. Removal of Fmoc group and rinsing is carried out, as described in the General methods.

Stage 5

(4S)-MeHex-D-Val-Thr(tBu)-Val-D-Val-D-Pro-Orn(Boc)-D-allo-Ile-D-allo-Thr(Val-Z-Dhb-Phe-Alloc)-D-allo-Ile-D-Val-O-TrtCl-resin

Alloc-group is removed using Pd(PPh3)4(58 mg, 0.05 mmol, 0.1 EQ.) in the presence of PhSiH3(617 μl, 5 mmol who, 10 equiv.) in the atmosphere of Ar and Alloc-Phe-Z-Dhb-HE (666 mg, 2 mmol, 4 equiv.) and HOAt (273 mg, 2 mmol, 4 equiv.) dissolved in DMF (1.25 ml) and added to peptidyl the resin, then add DIPCDI (310 μl, 2 mmol, 4 equiv.) and the mixture is stirred for 5 hours, after which the ninhydrin test is negative. After washings with DMF and CH2Cl2an aliquot peptidyl resin is treated with TFA-H2O (1:99) for 1 min and the product is characterized using MALDI-TOF-MS.

Calculated for C88H146N14O21: 1735,08.

Found: m/z 1758,67 [M+Na]+, 1774,62 [M+K]+.

Stage 6

(4S)-MeHex-D-Val-Thr(tBu)-Val-D-Val-D-Pro-Orn(Boc)-D-allo-Ile-D-allo-Thr(Val-Z-Dhb-Phe-N)-D-allo-Ile-D-Val-HE

After washings with DMF and CH2Cl2Alloc-group is removed using Pd(PPh3)4(58 mg, 0.05 mmol, 0.1 EQ.) in the presence of PhSiH3(617 μl, 5 mmol, 10 EQ.) in an atmosphere of Ar. The protected peptide otscheplaut from the resin using TFA-CH2Cl2(1:99) (5×30 sec). The filtrate is collected in N2O (4 ml) and N2About partially removed in a rotary evaporator. Then add TSA to dissolve the solids, which appears during the removal of the N2Oh, and the solution lyophilizer with getting 639 mg (387 mmol, yield 77%) specified in the connection header with the degree of purity >95%, as checked by HPLC (condition R, column C, tR=10,5 min).

Stadia

(4S)-MeHex-D-Val-Thr-Val-D-Val-D-Pro-Orn-D-allo-Ile-cyclo(D-allo-Thr-D-allo-Ile-D-Val-Phe-Z-Dhb-Val)

The protected peptide (stage 6) (639 mg, 387 μmol) dissolved in CH2Cl2(390 ml, 1 mm) and added HOBt (237 mg, 1.55 mmol)dissolved in minimum amount of DMF to dissolve HOBt add DIPEA (203 μl, of 1.16 mmol, 3 EQ.) and DIPCDI (240 μl, 1.55 mmol, 4 EQ.). The mixture is left to mix for 1 hour, then the course of the cyclization stage control using HPLC. The solvent is removed by evaporation under reduced pressure. The protected cyclic peptide was dissolved in TFA-H2O (19:1, 85 ml) and the mixture allowed to mix for 1 hour. The solvent is removed by evaporation under reduced pressure, added dioxane (30 ml) and the solvent is removed by evaporation under reduced pressure process (repeat three times), then add H2O (40 ml) and lyophilizers. The crude product is purified by HPLC (Kromasil C8, 5 μm, 205×50 mm), isocratic 44%acetonitrile (+0.05% of TFA) in water (+0.05% of TFA), 55 ml/h, the detection was performed at 220 nm, to obtain specified in the header of the product (192 mg, 0.13 mmol, yield 26%, 92.3 per cent).

MALDI-TOF-MS calculated for C75H124N14O16: 1476,93.

Found: m/z 1500,12 [M+Na]+, 1515,97 [M+K]+.

1H-NMR spectrum of the compound (2.5 mm, 500 MHz, H2O-D2O (9:1)) are presented in table II.

Table II
BalanceN-HOther
(Z)-Dhb9,59 (C)-6,63 (sq, J=7,5 Hz)1,19(d,γ-CH3)
D-allo-Ile 18,82 (d, J=9.0 Hz)4,421,871,25, 1,09, 0,82 (γ-CH2, γ-CH3, δ-CH3)
L-Pheis 8.75 (d, J=5,5 Hz)4,63is 3.08 (m)7,31 (2H Ar, t), 7,25 (3H Ar, d)
D-allo-Thr8,67(d, J=9.0 Hz)with 4.64of 5.05 (m)1-21 (γ-CH3)
D-Val 38,13(d, J=7.5 Hz)4,332,010,90 (2γ-CH3)
L-Orn8,29 (d, J=7.5 Hz)or 4.31of 1.66 (2H) 1,88 (γ-CH2), 2,96 (user. s, δ-CH2), 7,56 (ε-NH3+)
D-al.lo-Ile 27,92 (d)4,181,801,25, 1,09, 0,81 (γ-CH2, γ-CH3, δ-CH3)
D-Val 58,01 (d)4,082,070,87 (2γ-CH3)
L-Thr8,19 (d, J=7.5 Hz)4,294,14 (m)1,13 (γ-CH3)
D-Val 27,89 (d, J=7.5 Hz)4,322,110,78 (γ-CH3)
L-Val 48,04 (d)4,102,070,90 (2γ-CH3)
L-Val 17,19 (d, J=9.0 Hz)as 4.021,520,75 (γ-CH3), of 0.65 (d, γ-CH3)
D-Pro-4,36 2,23, 1,99 (m, β-CH2), 1-85 (m, γ-CH2), 3,83 (1H, m, δ-CH2), to 3.64 (1H, m, δ-CH2)
4(S)-MeHex-of 2.26 (2H)1,57 (β-CH2), 1-26, 1,10, 1,33, of 0.79 (δ-CH2, δ-CH3, γ-CH, ε-CH3)

Example 2

The analogs described in table III, synthesized using experimental techniques, as described in example 1, except for the stage indicated in column (stage), where the residue(s) (A) substituted(s) other(s) balance(s) (In) or removed (false).

Table III

Example 3

5-MeHex-D-Val-Thr-Val-D-Val-D-Pro-Orn-D-allo-Ile-cyclo[D-allo-Thr-D-alo-Ile-D-Val-hCh-(Z)-Dhb-Val](compound 63)

Experimental procedures as described in example 1 except that in stage 4 (4S)-MeHex substituted 5-MeHex and stage 5 is carried out in accordance with the following experimental method:

Alloc-group is removed from 5-MeHex-D-Val-Thr(tBu)-Val-D-Val-D-Pro-Orn(Boc)-D-allo-Ile-D-alo-Thr(Val-Alloc)-D-allo-Ile-D-Val-O-2-chlorotrityl-Ps (250 mg, initial load = 0.5 mmol/g resin) using, as mentioned above, Pd(PPh3)4in p outstay PhSiH 3in an atmosphere of Ar. Fmoc-Thr-OH (free hydroxyl group) (213,2 mg, 0,63 mmol, 5 EQ.) and Fmoc-hh-OH (254,0 mg, 0,63 mmol, 5 EQ.) successively added to the above peptidyl resin using DIPCDI (96.8 mg, 0,63 mmol, 5 EQ.) and HOBt (85 mg, 0,63 mmol, 5 EQ.) in DMF. After extensive washings with DMF (5×30 sec) peptidyl resin is treated with EDC·HCl (479 mg, 2.5 mmol, 20 equiv.) CuCl (184 mg, 1.5 mmol, 12 EQ.) in CH2Cl2-DMF (1:9) for 6 days. After extensive washes with DMF, CH2Cl2and DMF followed by the experimental protocols described in example 1 to obtain KF-equivalent.

Example 4

The analogs described in table IV, synthesized using experimental techniques, as described in example 1 except that in stage 4 (4S)-MeHex substituted 5-MeHex; stage 5 were carried out as described in example 3, but including Fmoc-h-OH instead of Fmoc-hh-OH, and at the stage shown in column (stage), remainder(s) (A) substituted(s) other(s) balance(s) (In) or removed(s) (no). In the case of analogues 64-66 not carry out the dehydration reaction, because the analogues gidrirovanny.

Table IV

Example 5

The analogs described in table V, synthesized using experimental techniques, as described in example 1, except that the stage 5 were carried out as described in example 3, and at the stage specified in number of the NECS (stage), the residue(s) (A) substituted(s) other(s) balance(s) (In). In addition, prior to solid-phase dehydration Fmoc group is removed as described in the General methods, and then the amino group protected in the form Alloc by interacting with Alloc-OSu (5 EQ.) in the presence of DIPEA (5 EQ.), using DMF as solvent (2 hours).

Table V

Example 6

[N(Me)2N'(Me)2-Arg8]-kahalalide F(compound 89)

DIPEA (1,73 μl, 10,17 mmol) and HATU (3,86 mg, 10,15 mmol) are added to kahalalide F (10.0 mg, 6,76 mmol)dissolved in DMF (5 ml). Interaction is monitored by HPLC and after 4 hours, the DMF is removed under reduced pressure and the residue dissolved in CH3CN-H2O-AcOH (4,5:4,5:1, 20 ml), lyophilizer and purified by prepreparation HPLC with obtaining specified in the header of the analogue of (4.5 mg, yield 40%).

Example 7

[N(Me,Ph),N'(Me)2-Arg8]-kahalalide F(compound 90)

Experimental procedures as described in example 6, but HAPyU (4,87 mg, 10,15 mmol) is used instead of HATU, getting to 1.63 mg, 12%.

Example 8

[N(CH2)4N'(Me)2-Arg8]-kahalalide F(compound 91)

Experimental procedures as described in example 6, but M2And (4,12 mg, 10,14 mmol) is used instead of HATU, gaining 5.2 mg, 46%.

Example 9

[N(CH2)4N'(CH2)4-Arg8]-kahalalide F(compound 92A)and [Nδ(N(CH2)4N'(CH2)4-UCP8]-kahalalide F(connection 92b)

Experimental procedures as described in example 6, but BTFFH (containing approximately 50% admixture of 1,1'-(permatile)dipyridine) (3,16 mg, 10,16 mmol) is used instead of HATU. Get two products, 92A and 92b, and the two cannot be separated (4.0 mg of the mixture in proportion 1:1, 35,6%).

HPLC (condition A, column (A), tR: 20,8 min (92b) with the release of 45% and tRis 20.9 min (92A) with the release of 46%.

EM (MALDI-TOF, m/z): (92A) calculated 1627,05; found 1630,8 [M+H]+; 1652,6 [M+Na]+. (92b) calculated 1629,06; found 1632,7 [M+H]+; 1670,6 [M+K]+.

Example 10

[Nε(Me)3-Lys8,(4S)-MeHex14]-KF(compound 93)

DIPEA (10 μl, 58.8 mmol) and MeI (6 μl, 0,100 mmol) are added to [Lys8,(4S)-MeHex14]-kahalalide F (compound 30) (5.0 mg, 3.35 mmol)dissolved in DMF/DCM (1:1, 5 ml). Interaction is monitored by HPLC and after 12 hours the solvent is removed under reduced pressure and the residue dissolved in CH3CN-H2O-AcOH (4,5:4,5:1, 20 ml), lyophilizer and purified by prepreparation HPLC with obtaining specified in the header of the analogue of (2.2 mg, yield 44%).

Example 11

[Thr(OTfa)12,4(S-MeHex 14]-kahalalide F(compound 94)

[4(S)-MeHex14]-kahalalide F (compound 1; 10.0 mg, 6.7 μmol) is dissolved in TFA/DCM (1:1, 20 ml) and left to mix for 3 days at room temperature. Then the solvent is removed under reduced pressure and the residue is dissolved in H2O-CH3CN (1:9) and immediately cleaned with a minimum of time during which the sample is dissolved in H2O. the Corresponding fraction is specified in the header analog collected in a round bottom flask immersed in liquid nitrogen, and lyophilizer (2.5 mg, yield 25%).

Example 12

[Orn(NδTfa)8,Thr(OTfa)12,4(S)-MeHex14]-kahalalide F(compound 95)

[(4S)-MeHex14]-KF (compound 1; 10 mg, 6.7 mmol) dissolved in DCM (8 ml), added TFAA (18,9 μl, 134 μmol) and DIPEA (22,8 μl, 134 mmol) and leave for interaction within 12 hours. Then the solvent is removed under reduced pressure, dissolved again in H2O-CH3CN (1:1) and immediately clean (2.0 mg, yield 20%).

Example 13

[Orn(NδTfa)8,4(S)-MeHex14]-kahalalide F(compound 96)

Experimental procedures as described in example 12, but before clearing the analogue dissolved in H2O-CH3CN (1:1)leave the solution for 2 hours and purified (4.3 mg, yield 43%).

Example 14

[Thr(OTfa)12,Lit(OTfa)14]-Kakha alid F (compound 97)

Experimental procedures as described in example 1 except that in stage 4 (4S)-MeHex replaced Lit-OH and on stage 7 of the cyclic peptide was dissolved in TFA/DCM (1:1, 20 ml) and left to mix for 3 days at room temperature. After this time the solvent is removed under reduced pressure from an aliquot (10%) and the solid residue was dissolved in H2O-CH3CN (1:9) and immediately cleaned with a minimum of time during which the sample is dissolved in N2O. the Fractions corresponding to the headline analogue, is collected in a round bottom flask immersed in liquid nitrogen, and lyophilizer (5.6 mg, yield 6% of the original resin).

Example 15

[no5-MeHex14-N-(Hep)2-D-Val13]-kahalalide F(compound 98)

Experimental procedures as described in example 1 except that in stage 4 of heptaldehyde (60,65 μl, 0.75 mmol, 5 equiv.) dissolved in DMF-Asón (99:1, 2 ml), add N-D-Val-Thr(tBu)-Val-D-Val-D-Pro-Orn(Boc)-D-allo-Ile-D-alo-Thr(Val-Alloc)-D-allo-Ile-D-Val-O-2-chlorotrityl-Ps (300 mg, the initial load = 0.5 mmol/g resin) and after 5 min add NaBH3CN (28,28 ml, 0.45 mmol, 3 EQ.), dissolved in DMF-Asón (99:1, 1 ml), and the mixture is left to interact for 2 hours. This process is repeated two more times for monitoring interaction using HPLC.

When is EP 16

[Orn(Biot)8]-kahalalide F(compound 99)

Kahalalide F (150,0 mg, 94,3 mmol), d-Biotin (37,0 mg, 151,5 mmol) and HATU (114,0 mg, 299,8 μmol) dissolved in anhydrous DCM (6.0 ml) in an atmosphere of Ar and added NMM (58 μl, 524,0 Microm). The mixture is left to mix for 20 hours. Then the solvent is removed under reduced pressure and the residue is dissolved in Meon and clean. Fractions corresponding to the headline analogue, lyophilizer (74,0 mg, yield 43%).

Characteristic analogues kahalalide F

The characteristics shown in table VI and table VII. EAT in table VI corresponds to MALDI-TOF, and the exact mass calculated using Chemwind 6,0, EAT in table VII corresponds chemical ionization at atmospheric pressure (ARS) with the formation of positive ions.

Table VI

Table VII

Example 17

Biological activity

The bioactivity of the compounds according to this invention is demonstrated by the results in the following tables, obtained in accordance with methods described in the following is the Braz.

The conclusion of this analysis is the cessation of growth, "in vitro" tumor cell cultures by continuous processing of the cells studied sample.

Cell line

NameNoATCCRodClothFeatures
A-549CCL-185peopleeasycarcinoma of the lung “NSCL”
SK-MEL-28HTB-72peoplemelanomamalignant melanoma
HT-29NTV-38peoplethe colonadenocarcinoma of the colon
LoVoCCL-229peoplethe colonadenocarcinoma of the colon
LoVo-Dox peoplethe colonadenocarcinoma of the colon (MDR)
DU-145HTB-81peopleprostateprostate cancer, pandrogeny receptors
LNCaPCRL-1740peopleprostateadenocarcinoma of the prostate with the androgen receptor
SK-BR-3NTV-30peoplemammary glandadenocarcinoma breast cancer, Her2/neu+
(pleural effusion)
IGROV-1peopleovaryadenocarcinoma of ovary
IGROV-ETpeopleovaryadenocarcinoma of ovary, defined as ET-743 resistant cells
SK-OV-3NTV-77people ovaryadenocarcinoma of ovary malignant ascites)
HeLaCCL-2peopleneckcervical epitelialna carcinoma
HeLa-LCCL-3peopleneckcervical epitelialna carcinoma, characterized as aplidin-resistant cells
K-562CCL-243peoplebone marrowchronic myelogenous leukemia
PANC-1CRL-1469peoplepancreasepitelialna carcinoma of the pancreas
NMES-1peopleendothelium

Colorimetric analysis type using reaction sulforhodamine In (SRB) adapted to quantify the growth and viability of cells [following methods is, described Philip Skehan and others (1990)New colorimetric cytotoxicity assay for anticancer drug screening, J. Natl. Cancer Inst., 82:1107-1112].

In this type of analysis using 96-well microplates for cell culture having a diameter of 9 mm (Faircloth, 1988; Mosmann, 1983). Most of the cell lines were obtained from American type culture collection (ATSS), originating from different types of human cancer.

Cells were kept in RPMI medium 1640 with 10% fetal calf serum, supplemented with 0.1 g/l penicillin and 0.1 g/l streptomycin sulfate, and then incubated at 37aboutC, 5% CO2and 98% humidity. For experiments cells were collected from subconfluent cultures using trypsin and re-suspended in fresh medium before cultivation.

Cells were planted in 96-well titration microplates, 5×103cells per well, in the aliquot Wednesday to 195 μl, and left them to attach to the tablet surface by cultivation free from drugs environment for 18 hours. Then, the samples were added to 5 µl aliquot in the range from 10 to 10-8μg/ml, dissolved in DMSO:EtOH:PBS (0,5:0,5:99). After 48 hours of exposure to anti-tumor effect was measured using the SRB methodology: the cells were fixed by adding 50 μl of cold 50% (wt./about.) trichloroacetic acid (TCA) and incubated for 60 min at 4aboutC. the Tablets were washed of Dionis is trated with water and dried. Added 100 μl of SRB solution (0.4 wt./vol.% in 1% acetic acid) to each well in the microtiter tablet, and incubated for 10 min at room temperature. Unbound SRB was removed by washing with 1% acetic acid. The tablets were dried in air and associated dye was solubilizers with Tris-buffer. Optical density was read on an automatic spectrophotometric tablet reader at a single wavelength of 490 nm.

Mean values ± standard deviation of the data was calculated from three holes. Some parameters of cellular responses can be calculated: GI = growth inhibition, TGI = total growth inhibition (cytostatic effect) and LC = lysis of the cells (cytotoxic effect).

Table VIII-IX illustrate data on the biological activity of the compounds according to the present invention.

Table VIII

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

T the blitz VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table VIII (PR is the continuation)

Data on the activity (mol.)

Table VIII (continued)

Data on the activity (mol.)

Table IX

Data on the activity (mol.)

1. The connection is based on the structure kahalalide F formulas (1)

denoted KF, and selected from the group consisting of [D-Ser6]-KF, [Glu8]-KF, [Lys8]-KF, [Val12]-KF,
[hCh11]-KF, [D-Cha4], D-Cha5D-Cha7]-KF, [Icos14]-KF, [(c/t)-4-Me-cHexa14]-KF, [Und14]-KF, [Oct14]-KF, [p-MeBza14]-KF, [Bza14]-KF, [R-CF3Bza14]-KF, [Pipe14]-KF, [R-CF3Cinn14]-KF, [p-CF3PhAcl4]-KF, [Pfh14]-KF, [6,6-dFHep14]-KF, [Lys8], (4S)-MeHexl4]-KF, [Gly13]-KF, [D-Ala13]-KF, [D-Leu13]-KF, [D-Phe13]-KF, [D-Pro13]-KF, [Val13]-KF, [D-Glu13]-KF, [D-Gln13]-KF, [D-Thr13]-KF, [Phe11]-KF, [Leu11]-KF, [Orn11]-KF, [Ala12, noD-Val13]-KF, [Gly12, noD-Val13]-KF, [Leu12, noD-Val13]-KF, [Glu12, noD-Val13]-KF, [D-Tic9, (4S)-MeHex14]-KF, [hCh3]-KF, [Dha2]-KF, [Trp3]-KF, [Phe(3,4-Cl2)3, (4S)-MeHex14]-KF, [Phe(F5)3, (4S)-MeHexl4]-KF, [Phe(4-I)3, (4S)-MeHexl4]-KF, [Phe(4-NO2)3, (4S)-MeHexl4]-KF, [Phe(4-F)3, (4S)-MeHex1 ]-KF, [Tyr(Me)3, (4S)-MeHex14]-KF, [Thi3, (4S)-MeHex14]-KF, [Tyr3, (4S)-MeHex14]-KF, [NMePhe3, (4S)-MeHex14]-KF, [Phe-(2-Cl)3]-KF, [Phe(3-Cl)3]-KF, [Phe(4-Cl)3]-KF, [Phe(3,4-F2)3]-KF, [Nal3]-KF, [Bip3]-KF, [Phe(3,4-Cl2)3p-CF3Cinn14]-KF, [N(Me)2N'(Me)2-Arg8]-KF, [N(Me,Ph),N'(Me)2-Arg8]-KF, [N(CH2)4N'(Me)2-Arg8]-KF, [N(CH2)4N'(CH2)4-Arg8]-KF, [Nδ(CHN(CH2)4-N(CH2)4)-Orn8]-KF, [Nε(Me)3-Lys8, (4S)-MeHex14]-KF, [Thr(OTFA)12, (4S)-MeHex14]-KF, [Orn(NδTFA)8, Thr(OTFA)12, (4S)-MeHex14]-KF, [Orn(NδTFA)8, (4S)-MeHex14]-KF, [Thr(OTFA)12, Lit(OTFA)14]-KF [Orn(Biot)8]-KF,
where the amino acid or group specified in square brackets, is a modification introduced in the structure kahalalide
F, or its pharmaceutically acceptable salt.

2. The connection is based on the structure kahalalide F formula (I):

denoted KF, and selected from the group consisting of [Gly12]-KF, [Ala12]-KF, [Leu12]-KF, [Phe12]-KF, [Orn12]-KF, [Pro12]-KF, [Gln12]-KF and [D-Val1]-KF, where the amino acid or group specified in square brackets, is a modification introduced in the structure kahalalide F, or its pharmaceutically acceptable salt.

3. Pharmaceutical composition, have the th antitumor activity, containing an effective amount of a compound according to any one of the preceding paragraphs and a pharmaceutically acceptable carrier, solvent or diluent.

4. The use of compounds according to claim 1 or 2 in getting medicines for cancer treatment.



 

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6 cl, 4 ex

FIELD: medicine.

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FIELD: chemistry, pharmacology.

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77 cl, 7 tbl, 17 dwg, 39 ex

FIELD: chemistry.

SUBSTANCE: invention concerns biochemistry, particularly biologically active peptides with stress-protection effect, which can be applied in medicine and pharmaceutics. The invention extends the range of safe substances stress resistance of an organism to include cyclopeptides I and II featuring the following aminoacid sequence: Cyclo(Gly1-Lys2-Val3-Leu4-Lys5-Lys6-Arg7-Arg8)n, where n = 2-3.

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5 tbl, 5 ex

FIELD: chemical technology.

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77 cl, 7 tbl, 16 dwg, 39 ex

FIELD: medicine, hormones, chemistry of peptides.

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EFFECT: valuable biological properties of compounds.

24 cl, 1 tbl, 2 ex

FIELD: organic chemistry, amino acids.

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EFFECT: valuable biological properties of compounds.

9 cl, 2 ex

FIELD: biochemistry.

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EFFECT: angiogenesis inhibiting cyclopeptides.

23 cl, 4 dwg, 2 tbl, 2 ex

The invention relates to compounds of formula (I) or (II)

or their pharmaceutically acceptable salts, where Y and Z each for each case independently represents a D - or L-natural or unnatural-amino acid; n in each case independently is 0 or 4, (I) provided that both n cannot simultaneously be 0; and 0 or 4 (II)

moreover, these amino acids (I) are chain: X1-X2-X3-X4where X1represents Tyr or Trp, which may be protected by a BOC group; X2represents D-Trp; X3represents Lys, which may be protected by a BOC group; X4is a Nal, Tyr or Thr; m is 0; a represents N or R1b means HE or OR1; (II) X1is a natural or non-natural D - or L-isomer of Phe, Trp or Tyr, where in the case when X1is Tyr, an aromatic ring in its side chain optionally substituted by R6; X2is a D - or L-isomer of Trp; X3represents Lys; X4represents Opticheskie ring, disposed in its side chain may be optionally substituted by R6or in the case when X4is either Ser or Thr, the oxygen atom located in its side chain, optionally may be substituted by one or more R1

FIELD: organic chemistry, antibiotics, medicine, oncology.

SUBSTANCE: invention relates to novel derivatives of aplidine possessing anti-tumor activity and represented by the general formula:

EFFECT: valuable medicinal property of compound, improved methods of synthesis and preparing.

28 cl, 3 tbl, 121 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and is a peptide which induces killer T cells ex vivo and which has an amino acid sequence as shown in one SEQ ID NOS: from 1 to 3. The disclosed peptide is used in an ex vivo agent for inducing anti-tumour immunity, in an ex vivo agent for inducing antigen-presenting cells, in an ex vivo agent which induces tumour-reactive T cells, as well as in an ex vivo pharmaceutical agent when treating or preventing tumours. The invention also relates to an antibody against the said peptide.

EFFECT: disclosed agents enable identification of glypican-3-derivative peptide, which can bond with HLA-A2, and activation of human killer T cells in order to provide an immunotherapy agent which may be effective in approximately 40% Japanese patients suffering from certain types of malignant tumours, accompanied by high level of GPC3 expression.

7 cl, 4 dwg, 1 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a pyrimidine nucleoside compound of general formula (1) , in which one of X and Y is a cyano group and the other is hydrogen; R1 is hydrogen, (R3)(R4)(R5)Si- or a carbonyl group which includes an alkyl monosubstituted with an amino group; R2 is hydrogen or (R6)(R7)(R8)Si-, provided that at least one of R1 and R2 is not hydrogen; or R1 and R2 together form a 6-member cyclic group -Si(R9)(R10)-, where each of R9 and R10 is a straight or branched alkyl; R3, R4 and R5 denote a straight or branched alkyl optionally substituted alkoxy, or cycloalkyl; R6, R7 and R8 denote a straight or branched alkyl optionally substituted alkoxy, cycloalkyl or phenyl, or to pharmacologically acceptable salts thereof. The invention also relates to a range of specific compounds of formula (1) or to their pharmacologically acceptable salts: 5'-O-triisopropylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-diethylisopropylsilyl-2'-cyano-2,-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-dimethylthexylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-(dimethyl-n-octylsilyl)-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-dimethylthexylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-diethylisopropylsilyl -2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-(tert-butyldimethylsily)-2'-cyano-2'-desoxy-1-β-O-arabinofuranosylcytosine; 3'-O-triisopropylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-dimethylthexylsilyl-5'-O-(L-valyl)-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-(L-valyl)-3'-O-(tert-butyldimethylsilyl)-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; and 3'-O-cyclopropyl-diisopropylsilyl-2'-cyano-2'-desoxy-1-β-D- arabinofuranosylcytosine.

EFFECT: obtaining formula (1) compounds or their pharmacologically acceptable salts for preparing a medicinal agent for treating tumours.

9 cl, 20 tbl, 1 dwg, 73 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of formula (I)

, pharmaceutical compositions based on the said compounds, as well as methods of using said compounds in preparing medicinal agents.

EFFECT: obtaining compounds and a composition which can inhibit phosphatase cdc25, particularly phosphatase cdc25-C and can be particularly used for treating cancer.

12 cl, 56 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula I and their pharmaceutically acceptable salts. The disclosed compounds have inhibitory effect on CDK1 kinase. In formula I , R1 is hydrogen or R2-(X)n-; X is a lower alkylene or cyclic lower alkylene; R2 denotes ; where denotes phenyl; cycloalkyl containing 3-6 carbon atoms; 4-6-member heterocycloalkyl ring having 3-5 carbon atoms and 1-2 oxygen atoms; R5, R6 and R7 are independently selected from a group containing hydrogen or halide; R4 is hydrogen or -(O)k(CH2CH2O)y-R10; R19 is hydrogen; R20 is hydrogen or -C(O)-R11; R10 and R11 is a lower alkyl; n and k are equal to 0 or 1; y is an integer from 0 to 3.

EFFECT: obtaining a pharmaceutical composition with inhibitory effect on CDK1 kinase, containing one or more of the disclosed compounds.

15 cl, 10 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to compounds of formula I and their pharmaceutically acceptable salts and esters. The disclosed compounds have inhibitory effect on cyclin-dependant kinase. In formula I R1 denotes , R3 is selected from a group consisting of H, CO2R6, C(O)R6, SO2R6 and SO2NR5R6, R5 and R6 are each independently selected from a group which includes H and (lower)alkyl, R2 is phenyl which contains one, two or three substitutes independently selected from a group which includes halogen or -O-(lower)alkyl.

EFFECT: preparation of a pharmaceutical composition which contains an effective amount of a formula I compound as an active ingredient.

6 cl, 1 tbl, 22 ex

FIELD: medicine.

SUBSTANCE: group of inventions relates to experimental medicine and can be applied for amplification of apoptosis or cytolytic activity in cells of mammals. Method and application by invention include bringing mammalian cells in contact with effective quantity of Apo- 2-ligand receptors agonist and resting or unprocessed NK-cells, said agonist is selected from Apo-2-ligand polypeptide, agonistic DR5 antibody and agonistic DR4 antibody.

EFFECT: application of inventions allows amplification of apoptosis and cytolysis induction in tumour cells due to activation by Apo-2-ligand receptors agonist of resting NK-cells.

26 cl, 1 tbl, 18 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine, pharmaceutical industry and deals with novel medications used for treatment of dysplastic processes of the cervix and rectum mucosa. Medication for treatment of dysplastic processes of the cervix and rectum mucosa, in form of suppository is characterised by the following: it contains 3,3'-diindolylmethane, epigallocatechin-3-gallate, as well as catalyst of epigallocatechin-3-gallate inhibiting activity with respect to DNA-methyltransferases, representing cations Mg2+ in form of pharmaceutically acceptable magnesium salt, lipophilic base, which contains hard fat, polyvinylpyrrolidone and butylhydroxyanisol and/or butylhydroxytoluol.

EFFECT: medication has efficient impact in case of severe forms of dysplastic injuries.

4 cl, 3 dwg, 5 ex, 2 tbl

FIELD: chemistry.

SUBSTANCE: present invention relates to biotechnology and immunology. An antibody against angiopoietin-2 is proposed. Versions of the antibody are disclosed, which are produced by hybridome ATCC PTA-7258, ATCC PTA-7259, ATCC PTA-7260. The corresponding coding nucleic acid and expression vector are disclosed. A host cell which produces the antibody based on the said vector is described. The disclosed antibodies have Kd of the order of 10-10-10-12 M, for the antibody 3.19.3 (from ATCC PTA-7260) IC50=99 nM. The said antibody properties can be used in treating human tumours.

EFFECT: design of a method of treating pathological angiogenesis based on an antibody and use of the antibody to prepare a medicinal agent for treating pathological angiogenesis.

33 cl, 18 dwg, 18 tbl, 24 ex

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